Condensed heterocyclic pyrazole derivatives as kinase inhibitors

ABSTRACT

Bicyclo-pyrazole compound of formula (I) and pharmaceutically acceptable salts thereof, as defined in the specification, processes for their preparation, combinatorial libraries comprising a plurality of them and pharmaceutical compositions thereof, are herewith disclosed: the compounds of the invention are useful, in therapy, as protein kinase inhibitors, for instance in the treatment of cancer.

CROSS REFERENCE TO RELATED APPLICATION

This application is a continuation of application Ser. No. 10/522,250filed on Sep. 19, 2005, which is a 371 of PCT/EP2003/007531 filed onJul. 11, 2003, which claims the benefit of U.S. Provisional ApplicationNo. 60/398,121 filed on Jul. 25, 2002.

The present invention relates to novel condensed heterocyclic pyrazolederivatives, to a process for their preparation, to pharmaceuticalcompositions comprising them and to their use as therapeutic agents, inparticular in the treatment of diseases linked to disregulated proteinkinases.

The malfunctioning of protein kinases (Pks) is the hallmark of numerousdiseases. A large share of the oncogenes and proto-oncogenes involved inhuman cancers code for PKs. The enhanced activities of PKs are alsoimplicated in many non-malignant diseases, such as benign prostatehyperplasia, familial adenomatosis, polyposis, neuro-fibromatosis,psoriasis, vascular smooth cell proliferation associated withatherosclerosis, pulmonary fibrosis, arthritis glomerulonephritis andpost-surgical stenosis and restenosis. PKs are also implicated ininflammatory conditions and in the multiplication of viruses andparasites. PKs may also play a major role in the pathogenesis anddevelopment of neurodegenerative disorders.

For a general reference to PKs malfunctioning or disregulation see, forinstance, Current Opinion in Chemical Biology 1999, 3, 459-465.

Accordingly, there is the need in therapy of compounds active inmodulating disregulated protein kinases, and in particular endowed withprotein kinase inhibiting activity.

The present inventors have now discovered that novel condensedheterocyclic pyrazole derivatives, according to the present invention,are endowed with modulating disregulated protein kinase activity, andare thus useful in therapy in the treatment of diseases associated withdisregulated protein kinases.

The present invention, as first object, provides a bicyclo-pyrazolecompound of formula (I):

wherein

-   X is NR′, O, S, SO or SO₂;-   each of R and R₁, being the same or different, is independently a    hydrogen atom or an optionally substituted group selected from R′,    —COR′, —COOR′, —CONHR′, —CONR′R″, —SO₂R′, —SO₂NHR′ or —SO₂NR′R″;    wherein each of R′ and R″, being the same or different, is    independently selected from hydrogen or an optionally further    substituted straight or branched C₁-C₆ alkyl, heterocyclyl, aryl or    aryl-C₁-C₆ alkyl group;-   R₂ is an optionally substituted group selected from R′, —CH₂OR′ and    OR′, wherein R′ is as above defined; and the pharmaceutically    acceptable salts thereof.

The compounds of formula (I), object of the present invention, may haveasymmetric carbon atoms and may therefore exist both as individualoptical isomers and as racemic admixtures thereof. Accordingly, all ofthe possible single isomers, including optical and geometrical isomers,of the compounds of formula (I) and admixtures thereof are also objectof the invention. The present invention comprises also the metabolitesand the pharmaceutically acceptable bio-precursors, otherwise referredto as pro-drugs, of the compounds of formula (I).

As it will be appreciated, the unsubstituted nitrogen atom in thecondensed pyrazole ring of the compounds of the invention can rapidlyequilibrate, in solution, as admixture of both tautomers:

Accordingly, where only one tautomer (I) or (Ia) is herein indicated forthe compounds of the invention, the other one and any mixture thereof,are also within the scope of the present invention, unless specificallynoted otherwise.

As used herein and unless other specified, the term straight or branchedC₁-C₆ alkyl, either as such or as a moiety in an aryl-alkyl group, ismeant a group chosen for instance from methyl, ethyl, n.propyl,isopropyl, n.butyl, isobutyl, sec-butyl, tert-butyl, n.pentyl, n.hexyland the like. Preferably it is a straight or branched C₁-C₄ alkyl, e.g.methyl, ethyl, n.propyl, isopropyl, n.butyl, isobutyl, sec-butyl andtert-butyl.

An aryl group, either as such or as aryl-alkyl group, is for instance amono-, bi- or poly-either carbocyclic as well as heterocyclichydrocarbon, with preferably from 1 to 4 ring moieties, either fused orlinked to each other by single bonds, wherein at least one of thecarbocyclic or heterocyclic rings is aromatic.

Non limiting examples of aryl groups are phenyl, indanyl, biphenyl, α-or, β-naphthyl, fluorenyl, 9,10-dihydroanthracenyl, pyridyl, pyrazinylpyrimidinyl, pyridazinyl, indolyl, imidazolyl, imidazopyridyl,1,2-methylenedioxyphenyl, thiazolyl, isothiazolyl, pyrrolyl,pyrrolyl-phenyl, furyl, phenyl-furyl, benzotetrahydrofuranyl, oxazolyl,isoxazolyl, pyrazolyl, chromenyl, thienyl, benzothienyl isoindolinyl,benzoimidazolyl, tetrazolyl, tetrazolylphenyl, pyrrolidinyl-tetrazolyl,isoindolinyl-phenyl, quinolinyl, isoquinolinyl, 2,6-diphenyl-pyridyl,quinoxalinyl, pyrazinyl, phenyl-quinolinyl, benzofurazanyl,1,2,3-triazolyl, 1-phenyl-1,2,3-triazolyl, and the like.

Accordingly, the term aryl group may also refer to aromatic carbocyclicor heterocyclic rings, further fused or linked to non aromaticheterocyclic rings, typically 5 to 7 membered heterocycles.

With the term 5 to 7 membered heterocycle, hence encompassing aromaticheterocycles also referred to as aryl groups, we also intend a saturatedor partially unsaturated 5 to 7 membered carbocycle, wherein one ormore, e.g. 1 to 3 carbon atoms, are replaced by heteroatoms such asnitrogen, oxygen and sulphur.

Examples of 5 to 7 membered heterocycles, optionally benzocondensed orfurther substituted, are 1,3-dioxolane, pyran, pyrrolidine, pyrroline,imidazolidine, pyrazolidine, pyrazoline, piperidine, piperazine,morpholine, tetrahydrofuran, azabicyclononane and the like.

According to the above meanings provided to R, R₁, R₂ R′ and R″, any ofthe above groups may be further optionally substituted, in any of theirfree positions, by one or more groups, for instance 1 to 6 groups,selected from: halogen, nitro, oxo groups (═O), carboxy, cyano, alkyl,perfluorinated alkyl alkenyl, alkynyl, cycloalkyl, aryl, heterocyclyl,amino groups and derivatives thereof such as, for instance, alkylamino,dialkylamino, arylamino, diarylamino, ureido, alkylureido or arylureido;carbonylamino groups and derivatives thereof such as, for instance,formylamino, alkylcarbonylamino, alkenylcarbonylamino,arylcarbonylamino, alkoxycarbonylamino, sulfonamido, alkylsulfonamidoand arylsulfonamido, hydroxy groups and derivatives thereof such as, forinstance, alkoxy, aryloxy, alkylcarbonyloxy, arylcarbonyloxy,cycloalkenyloxy or alkylideneaminooxy, carbonyl groups and derivativesthereof such as, for instance, alkylcarbonyl, arylcarbonyl,alkoxycarbonyl, aryloxycarbonyl, cycloalkyloxycarbonyl, aminocarbonyl,alkylaminocarbonyl, dialkylaminocarbonyl; sulfurated derivatives suchas, for instance, alkylthio, arylthio, alkylsulphonyl, arylsulphonyl,alkylsulphinyl, arylsulphinyl, arylsulphonyloxy, aminosulfonyl,alkylaminosulphonyl or dialkylaminosulphonyl. In their turn, wheneverappropriate, each of the above substituents may be further substitutedby one or more of the aforementioned groups.

A halogen atom is for instance fluorine, chlorine, bromine or iodine.

Any alkyl, alkenyl, alkynyl or alkoxy group can be either a branched orstraight moiety.

An alkyl group or moiety is, for instance, a C₁-C₆ alkyl group, inparticular a C₁-C₄ alkyl group as exemplified above.

An alkoxy group is for instance a C₁₋₆ alkoxy group, in particular aC₁-C₄ alkoxy group such as methoxy, ethoxy, propoxy, isopropoxy, butoxy,sec-butoxy or tert-butoxy.

An alkenyl or alkynyl group is for instance a C₂-C₆ alkenyl or alkynylchain, in particular a C₂-C₄ alkenyl or alkynyl chain, preferably avinyl, allyl or ethynyl group, respectively.

An aryl or heterocyclyl group or moiety is, for instance, an aryl orheterocyclyl group or moiety, as defined above.

A cycloalkyl group is, for instance, a C₃-C₆ cycloalkyl group such ascyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

Pharmaceutically acceptable salts of the compounds of formula (I) arethe acid addition salts with inorganic or organic acids, e.g. nitric,hydrochloric, hydrobromic, sulphuric, perchloric, phosphoric, acetic,trifluoroacetic, propionic, glycolic, lactic, oxalic, malonic, malic,maleic, tartaric, citric, benzoic, cinnamic, mandelic, methanesulphonic,isethionic and salicylic acid, as well as the salts with inorganic ororganic bases, e.g. alkali or alkaline earth metals, especially sodium,potassium, calcium or magnesium hydroxides, carbonates or bicarbonates,acyclic or cyclic amines, preferably methylamine, ethylamine,diethylamine, triethylamine or piperidine.

Preferred compounds of the invention are the compounds of formula (I)wherein X is S or O; R is —CONHR′; R₁ is —COR′, —CONHR′, —CONR′R″,—SO₂NHR′ or —SO₂NR′R″, wherein each of R′ and R″, being the same ordifferent, is selected from hydrogen or an optionally substitutedstraight or branched C₁-C₆ alkyl, aryl and aryl-C₁-C₆ alkyl group; R₂ ishydrogen; and the pharmaceutically acceptable salts thereof.

Even more preferred, are the compounds of the invention of formula (I)wherein X is S; R is —CONHR′; R₁ is —CONHR′ or —CONR′R″, wherein each ofR′ and R″, being the same or different, is selected from hydrogen or anoptionally substituted straight or branched C₁-C₆ alkyl, aryl oraryl-C₁-C₆ alkyl group; R₂ is hydrogen; and the pharmaceuticallyacceptable salt thereof.

For a general reference to any specific compound of formula (I) of theinvention, optionally in the form of a pharmaceutically acceptable salt,see the experimental section and claims.

As formerly indicated, as an additional object of the invention herewithprovided are the compounds of formula (I), or the pharmaceuticallyacceptable salts thereof, for use as a medicament.

In addition, the present invention also provides the use of thecompounds of formula (I), or the pharmaceutically acceptable saltsthereof, in the manufacture of a medicament for treating a patientsuffering from a disease caused by and/or associated with an altered(disregulated) protein kinase activity.

The present invention thus provides a method for treating a mammal,including humans, suffering from a disease caused by and/or associatedwith an altered (disregulated) protein kinase activity, which comprisesadministering to said mammal in need thereof a therapeutically effectiveamount of a bicyclo-pyrazole compound of formula (I):

wherein

-   X is NR′, O, S, SO or SO₂;-   each of R and R₁, being the same or different, is independently a    hydrogen atom or an optionally substituted group selected from R′,    —COR′, —COOR′, —CONHR′, CONR′R″, —SO₂R′, —SO₂NR′ or —SO₂NR′R″;    wherein each of R′ and R″, being the same or different, is    independently selected from hydrogen or an optionally further    substituted straight or branched C₁-C₆ alkyl, heterocyclyl, aryl or    aryl-C₁₋₆ alkyl group;-   R₂ is an optionally substituted group selected from R′, —CH₂OR′ and    OR′, wherein R′ is as above defined; and the pharmaceutically    acceptable salts thereof.

In a preferred embodiment, the disease caused by and/or associated withan altered (disregulated) protein kinase activity is selected from thegroup consisting of cancer, cell proliferative disorders, Alzheimer'sdisease, viral infections, autoimmuno diseases and neurodegenerativedisorders.

Specific preferred types of cancer that may be treated includecarcinoma, squamous cell carcinoma, hematopoietic tumors of myeloid orlymphoid lineage, tumors of mesenchymal origin, tumors of the centraland peripheral nervous system, melanoma, seminoma, teratocarcinoma,osteosarcoma, xeroderma pigmentosum, keratoxanthomas, thyroid follicularcancer and Kaposi's sarcoma.

In another preferred embodiment of the method for treating a mammaldescribed above, the cell proliferative disorder is selected from thegroup consisting of benign prostate hyperplasia, familial adenomatosispolyposis, neuro-fibromatosis, psoriasis, vascular smooth cellproliferation associated with atherosclerosis, pulmonary fibrosis,arthritis glomerulonephritis and post-surgical stenosis and restenosis.

In addition, the method for treating a mammal object of the presentinvention also provides tumor angiogenesis and metastasis inhibition.

A further object of the invention is also represented by the process forpreparing the compounds of formula (I) of the invention, and thepharmaceutically acceptable salts thereof, which process comprises:

-   a) reacting a compound of formula (II)

wherein R₁, R₂ and X are as defined above and R₃ is a lower alkyl group,with hydrazine or a hydrazine salt, so as to obtain a compound offormula (I) wherein R is hydrogen and R₁, R₂ and X are as defined above;and, if desired,

-   b) converting the thus obtained compound of formula (I) into another    compound of formula (I) wherein R is other than a hydrogen atom;    and/or, if desired, converting the compound of formula (I) into a    pharmaceutically acceptable salt thereof and/or the salt of a    compound of formula (I) into a free compound thereof.

The above process is an analogy process, which can be carried outaccording to known methods in the art.

From the above, it is clear to the person skilled in the art that if acompound of formula (I), prepared according to the above process, isobtained as an admixture of isomers, their separation into the singleisomers of formula (I), carried out according to conventionaltechniques, is still within the scope of the present invention.

Likewise, salification of a compound of formula (I) or conversion of acorresponding salt thereof into the free compound (I), according to step(b) of the process, can be easily carried out according to well-knownmethods in the art.

According to step a) of the process, the reaction between a compound offormula (II) and hydrazine or a hydrazine salt, for instance hydrazinedihydrochloride or hydrazine sulphate or acetate, can be carried out inthe presence of catalytic amounts of an acid such as hydrochloric,acetic or sulphuric acid, or in the presence of catalytic amounts of aLewis acid such as boron trifluoride dimethyl etherate, or even in thepresence of catalytic amounts of a strong base such us sodium methoxide.

The reaction is carried out in a suitable solvent such as, for instance,N,N′-dimethylformamide, tetrahydrofuran, 1,4-dioxane, acetonitrile,water, methanol or ethanol, at a temperature ranging from about roomtemperature to reflux and for a time varying from about 30 minutes toabout 18 hours.

According to a preferred embodiment, within the compounds of formula(II), R₃ is a straight or branched lower alkyl group, for instance aC₁-C₆ alkyl group and, even more preferably, a C₁-C₄ alkyl group.

Preferably, step a) is carried out by reaction of a compound of formula(II) and hydrazine hydrate in ethanol, at a temperature ranging fromroom temperature and refluxing temperature followed by cyclization ofthe intermediate 2-hydrazinothiophene thus obtained. Cyclization iscarried out at a temperature ranging from about 15° C. to about 50° C.in methanol or ethanol, and in the presence of catalytic amounts of amineral acid such as hydrochloric or sulphuric acid.

The thus obtained bicyclo-pyrazoles of formula (I) and wherein R ishydrogen, as per step a), can be easily converted into a variety ofderivatives of formula (I) having R other than a hydrogen atom, and/orinto salts thereof.

As set forth in step (b), any of these conversions may occur accordingto conventional techniques, by properly reacting the amino derivative(I) with alkylating, acylating, sulfonylating agents and the like.

In this respect it is worth noting that optional by-products, forinstance originating by the above reactions also occurring at thenitrogen pyrazole atom, may be obtained. As such, a subsequent step toisolate the desired compound of formula (I) is then required and carriedout according to the methods known in the art.

Optionally, compounds obtained by reacting the amino derivative (I) withacylating agents and wherein acylation occurred at both the amino groupand the nitrogen pyrazole atom, can be easily converted into thecorresponding compounds of formula (I) by selective hydrolysis of theacyl group at the pyrazole nitrogen atom.

Likewise, a compound of formula (I) obtained according to step a) above,can be first reacted with an opportune acylating agent to protect thepyrazole nitrogen atom, then reacted as per step b), and finallyde-protected at the pyrazole nitrogen atom.

Alternatively, a compound of formula (I) being obtained according tostep a), may be first supported onto a suitable resin and, then reactedas per step b) above followed by resin cleavage.

Therefore, object of the present invention is also a process forpreparing a compound of formula (I) and the pharmaceutically acceptablesalts thereof, which process comprises:

-   a) reacting a compound of formula (II) as above defined with    hydrazine or a hydrazine salt so as to obtain a compound of    formula (I) wherein R is a hydrogen atom and R₁, R₂ and X are as    defined above;-   a′) reacting the thus obtained compound of formula (I) with an alkyl    chlorocarbonate of formula (III)    R₃—O—CO—Cl  (III)    wherein R₃ is a lower allyl group, so as to obtain a compound of    formula (IV) being protected at the heterocyclic pyrazole nitrogen    atom:

-   b) converting the thus obtained compound of formula (IV) into a    compound of formula (V)

wherein R, being as defined above, is other than a hydrogen atom and X,R₁, R₂ and R₃ are as defined above; and

-   c) cleaving the protecting group under alkaline conditions so as to    obtain the desired compound of formula (I) and, if desired,    converting the thus obtained compound of formula (I) into another    compound of formula (I); and/or, if desired, converting the compound    of formula (I) into a pharmaceutically acceptable salt thereof    and/or the salt thereof into a free compound of formula (I).

In step a′), the reaction between the alkyl chlorocarbonate of formula(III) and the compound of formula (I), as therein defined, can becarried out in a suitable solvent such as, tetrahydrofuran,dichloromethane, chloroform, acetonitrile, toluene or a mixture thereof,at a temperature ranging from about −5° C. to about 35° C. and for atime varying from about 30 minutes to about 72 hours, in the presence ofan opportune proton scavenger such as triethylamine ordiisopropylethylamine.

After having carried out the conversion of a compound of formula (IV)into a compound of formula (V), as described below, the subsequentde-protection, in step c), is carried out under alkaline conditions byworking according to conventional techniques, for instance by treatmentwith aqueous sodium or potassium hydroxide in the presence of a suitableco-solvent such as methanol, ethanol, dimethylformamide, 1,4-dioxane, orby treatment with a tertiary amine such as triethylamine ordisopropylethylamine using an alcohol like methanol or ethanol as thesolvent.

Deprotection occurs at a temperature ranging from about 18° C. torefluxing temperature of the solvent, for a time varying from about 30minutes to about 72 hours.

A further object of the invention is also a process for preparing acompound of formula (I) and the pharmaceutically acceptable saltsthereof, in which X, R₁ and R₂ are as defined above and R is anoptionally substituted group selected from —COR′ or —COOR′, whichprocess comprises:

-   a″) reacting a compound of formula (I) as obtained in the above    step (a) and wherein R is a hydrogen atom with an excess of a    chlorocarbonate or an acyl chloride derivative of formula (IIIa) or    (IIIb), respectively:    R′—O—CO—Cl  (IIIa)    R′—CO—Cl  (IIIb)    wherein R′ is as above defined, so as to obtain a compound of    formula (Va) or (Vb), respectively

-   c′) cleaving under alkaline conditions the compound of formula (Va)    or (Vb) so as to eliminate the protecting group at the pyrazole    nitrogen atom and thus obtaining the desired compound of formula (I)    and, if desired, converting the thus obtained compound of    formula (I) into another compound of formula (I); and/or, if    desired, converting the compound of formula (I) into a    pharmaceutically acceptable salt thereof and/or the salt thereof    into a free compound of formula (I).

In step a″), the reaction between the compound of formula (IIIa) or(IIIb) with the compound of formula (I), as therein defined, can becarried out in a suitable solvent such as, tetrahydrofuran,dichloromethane, chloroform, acetonitrile, toluene or a mixture thereof,at a temperature ranging from about −5° C. to about 35° C. and for atime varying from about 30 minutes to about 72 hours, in the presence ofan opportune proton scavenger such as triethylamine ordiisopropylethylamine.

The subsequent selective hydrolysis of the acyl group at the pyrazolenitrogen atom, in step c′), is carried out under alkaline conditions byworking according to conventional techniques, as formerly reported.

According to a further aspect of the process, a compound of formula (I)being obtained as per step a) above, is first supported onto a suitableresin and, then, reacted as per step b). Therefore an additional objectof the invention is also a process for preparing a compound of formula(I) and the pharmaceutically acceptable salts, which process comprises:

-   a′″) reacting a compound of formula (I) wherein R is a hydrogen atom    and R₁, R₂ and X are as above defined; with an isocyanate    polystyrenic resin of formula (VI)

so as to obtain a polystyrenemethyl urea; of formula (VII)

-   b) converting the thus obtained compound of formula (VII) into a    compound of formula (VIII)

wherein R, being as defined above, is other than a hydrogen atom; and

-   c″) cleaving under alkaline conditions the compound of    formula (VIII) so as to eliminate the resin and thus obtaining the    desired compound of formula (I) and, if desired, converting the thus    obtained compound of formula (I) into another compound of formula    (I); and/or, if desired, converting the compound of formula (I) into    a pharmaceutically acceptable salt thereof and/or the salt thereof    into a free compound of formula (I).

In step a′″), the reaction between the isocyanatomethyl polystyrenicresin of formula (VI) and the compound of formula (I), as thereindefined, can be carried out in a suitable solvent such as,N,N-dimethylformamide, dichloromethane, chloroform, acetonitrile,toluene or a mixture thereof, at a temperature ranging from about 5° C.to about 35° C. and for a time varying from about 30 minutes to about 72hours.

After having carried out the conversion of a compound of formula (VII)into a compound of formula (VIII), as formerly described, the subsequentcleavage of the resin, in step c″), is carried out under alkalineconditions by working according to conventional techniques. As anexample, aqueous sodium or potassium hydroxides in the presence of asuitable co-solvent such as methanol, ethanol, dimethylformamide,1,4-dioxane, or acetonitrile can be employed.

The conversion of a compound of formula (VII) into a compound of formula(VIII) can be carried out under mild operative conditions, attemperatures varying from about 5° C. to about 60° C. and for a timevarying from about 2 hours to about 7 days.

As far as step b) of the process is concerned, the optional conversionof a compound of formula (I) or, more preferably, of formulae (IV) or(VII), into the corresponding derivatives of formula (I), (V) or (VIII),respectively, and having R other than hydrogen, can be carried accordingto conventional techniques known in the art to alkylate, acylate orsulfonylate amino groups.

In particular, a compound of formula (I) wherein R, being other thanhydrogen, is selected from: R′, —COR′, —COOR′, —SO₂R′, —SO₂NHR′ and—SO₂NR′R″, wherein R′ and R″ have the values as given in formula (I)above; and R₁ and R₂ are as defined above, may be prepared by reacting acompound of formula (I), wherein R is hydrogen, with a correspondingcompound of formula (IX)R—Y  (IX)wherein R has the above reported meanings other than hydrogen, and Y isa suitable leaving group, preferably chlorine or bromine.

The above conditions apply, by analogy, either to the conversion of acompound of formula (IV) into a compound of formula (V), throughreaction with a derivative o formula (IX), as well as to the conversionof a compound of formula (VII) to the compound of formula (VIII),through reaction with a derivative o formula (IX).

The above reaction can be carried out according to conventionalprocedures known in the art for acylating, sulfonylating or alkylatingamino groups, for instance in the presence of a tertiary base, such astriethylamine, N,N-diisopropylethylamine or pyridine, in a suitablesolvent such as toluene, dichloromethane, chloroform, diethyl ether,tetrahydrofuran, acetonitrile, or N,N-dimethylformamide, at atemperature ranging from about −10° C. to reflux and for a time varyingfrom about 30 minutes to about 96 hours. From the foregoing, it is clearto the person skilled in the art that the preparation of the compoundsof formula (I) having R equal to —SO₂NR′R″ can be actually performed asabove described or, alternatively, by properly reacting a compound offormula (I) or (V) having R equal to —SO₂NHR′ with any suitablealkylating moiety, according to well known methodologies for preparingdi-substituted sulfonamides.

A compound of formula (I) wherein R is a —CONHR′ group, R′ has the abovereported meanings other than hydrogen, X, R₁ and R₂ are as abovedefined, may be prepared by reacting a compound of formula (I) having Requal to hydrogen, with a compound of formula (X)R′NCO  (X)wherein R′ is as above defined other than hydrogen.

The above conditions apply, by analogy, either to the conversion of acompound of formula (IV) into a compound of formula (V), throughreaction with a derivative o formula (X), as well as to the conversionof a compound of formula (VII) to the compound of formula (VIII),through reaction with a derivative o formula (X). Compounds of formula(I), (V) or (VIII) wherein R is a —CONHR′ group may be optionallyfurther reacted with a compound of formula (XI)R″—Y  (XI)wherein R″ is as above defined other than hydrogen and Y is a suitableleaving group, preferably chlorine or bromine, so as to obtain thecorresponding compounds of formulae (I), (V) or (VIII) wherein R is—CONR′R″, being R′ and R″ other than hydrogen atoms. The reactionbetween the above compounds (I), (IV) or (VII) with a compound offormula (X) can be carried out in the presence of a tertiary base, suchas triethylamine, N,N-diisopropylethylamine or pyridine, in a suitablesolvent, such as toluene, dichloromethane, chloroform, diethyl ether,tetrahydrofuran, acetonitrile, or N,N-dimethylformamide, at atemperature ranging from about −10° C. to reflux. The reaction time mayvary from about 30 minutes to about 72 hours.

The optional subsequent conversion of a compound of formula (I), (V) or(VIII) having R equal to —CONHR′ into a corresponding derivative havingR equal to —CONR′R″ is carried out according to conventional methodsused to prepare di-substituted ureido derivatives.

A compound of formula (I) or (V) wherein R is a —CONR′R″ group, R′ andR″ have the above reported meanings other than hydrogen, X, R₁ and R₂are as above defined, may be prepared by reacting a compound of formula(I) or (IV) having R equal to hydrogen, with 4-nitrophenylchloroformateand subsequently with a compound of formula (XII)R′R″NH  (XII)wherein R′ and R″, are as defined above other than hydrogen.

The reaction is carried out according to conventional methods used toprepare di-substituted ureido derivatives.

Analogously, a compound of formula (VIII) wherein R is a —CONR′R″ group,R′ and R″ have the above reported meanings other than hydrogen, X, R₁and R₂ are as above defined, may be prepared by reacting a compound offormula (VII), having R equal to hydrogen with4-nitrophenylchloroformate and, subsequently, with a compound of formula(XII) as defined above.

Alternatively, a compound of formula (I), or a compound of formula (IV),having R equal to hydrogen, may be reacted under reductive conditionswith a compound of formula (XIII)R′—CHO  (XIII)wherein R′, being as defined above is other than hydrogen, so as toobtain a corresponding compound of formula (I) or (V) wherein R is aR′CH₂— group and R′ being as defined above is other than hydrogen.

Analogously, a compound of formula (VII) heaving R equal to hydrogen maybe reacted under reductive conditions with a compound of formula (XIII)as defined above, so as to obtain a corresponding compound of formula(VIII) wherein R is a R′CH₂— group and R′ being as defined above isother than hydrogen.

The reaction is carried out in a suitable solvent such as, for instance,N,N-dimethylformamide, N,N-dimethylacetamide, chloroform,dichloromethane, tetrahydrofuran, or acetonitrile, optionally in thepresence of acetic acid, ethanol or methanol as co-solvents, at atemperature ranging from about −10° C. to reflux and for a time varyingfrom about 30 minutes to about 4 days.

Conventional reducing agents in the reaction medium are, for instance,sodium boron hydride, sodium triacethoxy boron hydride, and the like.

From the foregoing, it is clear to the person skilled in the art thanany of the above compounds of formula (I), (IV), (V), (VII) and (VIII)may be conveniently converted into other derivatives (I), (IV), (V),(VII) or (VIII) also by properly reacting functional groups other thanthe R group, extensively described above, according to conventionalsynthetic organic methods.

In a further example, the compounds of formula (I) or (VIII) wherein R₁is —COOMe can be hydrolized to the corresponding compounds of formula(I) or (VIII) wherein R₁ is —COOH, by treatment with a suitable base,for instance sodium or potassium hydroxide, according to conventionalsynthetic organic methods.

Any of the above compounds of formula (I) or (VIII) and where R₁ is a—COOH group can be easily converted into other derivatives (I) or (VIII)by properly reacting the carboxylic group according to conventionalsynthetic organic methods.

In particular, a compound of formula (I) or (VIII) wherein R₁ is a—CONR′R″ group, R′ and R″ have the above reported meanings other thanhydrogen, R is as above defined other than hydrogen, and X and R₂ are asabove defined, may be prepared by reacting a corresponding compound offormula (I) or (VIII), wherein R₁ is a —COOH group, with a condensingagent and, subsequently, with a compound of formula (XII)R′R″NH  (XII)wherein R′ and R″, are as defined above.

The reaction is carried out according to conventional methods used toprepare amides. Likewise, a compound of formula (I) or (VIII) wherein R₁is —COR′, R′ has the above reported meanings, R is other than hydrogen,and R₂ and X are as above defined, may be prepared by reacting acorresponding compound of formula (I) or (VIII), wherein R₁ is a Weinrebamido —CONCH₃OCH₃ group with a compound of formula (XIV)R′Li  (XIV)wherein R′ is as defined above other than hydrogen.

The reaction is carried out according to conventional methods used toprepare ketones. As an example, the reaction between the above compounds(I) or (VIII) with a compound of formula (XIV) can be carried out in asuitable solvent, such as tetrahydrofuran, toluene, diethyl ether,hexane, at a temperature ranging from about −78° C. to about 10° C. andfor a time varying from about 10 minutes to about 72 hours. A compoundof formula (I) or (V) wherein R₁ is a Weinreb amido CONCH₃OCH₃ group canbe obtained by known methods.

Alternatively a compound of formula (I) or (VIII) wherein R₁ is a —COOR′group, R′ has the above reported meanings other than hydrogen, R isother than hydrogen, and R₂ and X are as above defined, may be preparedby reacting a corresponding compound of formula (I) or (VIII), whereinR₁ is a —COOH group with a compound of formula (XV)R′OH  (XV)wherein R′ is as defined above other than hydrogen.

The reaction is carried out according to conventional methods used toprepare esters. Likewise, the preparation of the compounds of formula(V) or (VIII) having R₁ equal to —SO₂NR′R″ can be actually performed asdescribed below or, alternatively, by properly reacting a compound offormula (V) or (VIII) having R₁ equal to —SO₂NHR′ with any suitablealkylating moiety, according to well known methodologies for preparingdi-substituted sulfonamides.

From the foregoing, it is clear to the person skilled in the art thanany of the above compounds of formula (I) and (VIII) may be convenientlyconverted into other derivatives (I) or (VIII) also by properly reactingfunctional groups other than the R and R₁ groups, extensively describedabove, according to conventional synthetic organic methods.

As an example, a compound of formula (I) or (VIII) wherein X is SO, R isother than hydrogen, and R₁ and R₂ are as defined above, may beconveniently prepared by starting from a corresponding compound offormula (I) or (VIII) wherein X is S with an oxidizing agent accordingto conventional synthetic organic methods. The reaction can be carriedout in the presence of an oxidizing agent such as, for instance,boron-trifluoride diethyl etherate in the presence of MCPBA, hydrogenperoxide in the presence of TFA and the like, in a suitable solvent suchas dichloromethane, water, methanol, ethanol at a temperature rangingfrom about −10° C. to reflux and for a time varying from about 30minutes to about 48 hours.

In a further example, a compound of formula (I) or (VIII), wherein X isSO₂, R is other than hydrogen, and R₁ and R₂ are as defined above, maybe conveniently prepared by starting from a corresponding compound offormula (I) or (VIII) wherein X is S, with an oxidizing agent accordingto conventional synthetic organic methods. As an example, the reactioncan be carried out in the presence of an oxidizing agent, such as, forinstance, MCPBA, dimethyldioxirane, oxone, Mg monoperoxyphthalate, in asuitable solvent such as, dichloromethane, chloroform, acetone,acetonitrile, water, methanol, ethanol, at a temperature ranging fromabout −10° C. to reflux and for a time varying from about 30 minutes toabout 48 hours.

In a further example, a compound of formula (V) or (VIII) wherein X isNR′, R′ and R are other than hydrogen atoms, and R₁ and R₂ are asdefined above, may be conveniently prepared by staring from acorresponding compound of formula (V) or (VIII) wherein X is NH, throughreaction with a compound of formula (XVI)R′—Y  (XVI)wherein R′, being as defined above, is other than hydrogen and Y is asuitable leaving group, preferably chlorine or bromine. This latterreaction can be carried out in the presence of a base such as sodiumhydride, tBuOK, potassium carbonate, potassium hydroxide and the like,in a suitable solvent such as toluene, dichloromethane, chloroform,diethyl ether, tetrahydrofuran, acetonitrile, N,N-dimethylformamide ordimethylsulfoxyde, at a temperature ranging from about −10° C. toreflux.

From the foregoing, it is clear to the person skilled in the art thatany of the above compounds of formula (I), (V) and (VIII) may beconveniently converted into other derivatives (I), (V) or (VIII) also byproperly reacting functional groups other than the R, R₁ or X groups,extensively described above, according to conventional synthetic organicmethods.

As an example, the above compounds (I) or (VIII) having R₂ equal to—CH₂OH or —OH, R other than hydrogen, and R₁ and X as above defined, canbe reacted with a compound of formula (XVI)R′—Y  (XVI)wherein R′, being as defined above is other than hydrogen and Y is asuitable leaving group as defined above, so as to obtain thecorresponding compounds having R₂ as —CH₂OR′ or OR′ group.

This latter reaction can be carried out in the presence of a base suchas sodium hydride, N,N-diisopropylethylamine or pyridine, in a suitablesolvent, such as toluene, dichloromethane, chloroform, diethyl ether,tetrahydrofuran, acetonitrile, or N,N-dimethylformamide, at atemperature ranging from about −10° C. to reflux.

As it will be really appreciated by the person skilled in the art whenpreparing the compounds of formula (I) object of the invention, optionalfunctional groups within both the staring materials or the intermediatesthereof and which could give rise to unwanted side reactions, need to beproperly protected according to conventional techniques. Likewise, theconversion of these latter into the free deprotected compounds may becarried out according to known procedures.

All of the compounds of formula (II), (III), (VI), (IX), (X), (XI),(XII), (XIII), (XIV), (XV) and (XVI) according to the process object ofthe invention, comprehensive of any variant thereof, are known or can beobtained according to known methods.

In particular, the compounds of formula (III), (VI), (IX), (X), (XI),(XII), (XIII), (XIV), (XV), (XVI), and (XVII) are commercially availableor readily prepared according to conventional methods; the compounds offormula (II), instead, can be prepared by oxidising a compound offormula (XVII)

wherein R₁, R₂ and X are as above defined and R₃ is a lower alkyl group,according to conventional methods reported in the literature. As anexample, the reaction can be carried in the presence of an opportuneoxidizing agent such as, for instance, hydrogen peroxide,3-chloroperoxybenzoic acid, oxone, in a suitable solvent such as, forinstance, dichloromethane, DMF, acetone, toluene, acetonitrile,methanol, ethanol, water, acetic acid, at a temperature ranging fromabout −10° C. to reflux and for a time varying from about 30 minutes toabout 4 days.

Compounds of formula (XVII) wherein R₁, R₃ and X are as above definedand R₂ is a hydrogen atom can be prepared by working in analogy to theprocedure described by Wilson K. J. et al., in J. Bioorg. Med. Chem.Lett. 11 (2001), 915-918, starting from a compound of formula (XVIII)

The compounds of formula XVII wherein R₁, R₃ and X are as above definedand R₂ is other than a hydrogen atom can be prepared by treating acompound of formula (XVIII) with sodium nitrite, and by subsequentlyreacting the thus obtained-diazonium salt according to conventionalmethods in order to obtain the desired R₂ substituents.

Also the compounds of formula (XVIII) wherein R₁ is —COOMe, R₃ is amethyl group and X is S, can be prepared according to the aforementionedprocedures described by Wilson K. J. et al., by starting from a compoundof formula (XV).

The corresponding compounds of formula (XVIII) wherein R₁ is —COOR′,—SO₂R′, —SO₂NHR′ or SO₂NR′R″, and R₃ and X are as above defined, can bealso prepared analogously by using the aforementioned proceduresdescribed by Wilson K. J. et al.

A compound of formula (XVIII) wherein R₁ is —COOtBu, can be convenientlyprepared by the corresponding compound of formula (XVIII) wherein R₁ is—COOMe by using standard procedures well known in the art.

A compound of formula (XVIII) wherein R₁ is —COOR′, —SO₂R′, —SO₂NHR′ orSO₂NR′R″, and R₃ and X are as above defined, can be convenientlyprepared by reacting a compound of formula (XIX)

wherein R₁ and X are as defined above, with a compound of formula (XX)

wherein R₃ is as defined above.

This latter reaction can be carried out in the presence of a base, suchas triethylamine, N,N-diisopropylethylamine, pyridine or sodium hydridein a suitable solvent such as toluene, diethyl ether, tetrahydrofuran,acetonitrile, dimethyl sulfoxide or N,N-dimethylformamide, at atemperature ranging from about 10° C. to reflux and for a time varyingfrom about 30 minutes to about 96 hours.

In a further example, a compound of formula (XVIII) wherein R₁ is R′,and R₃ and X are as above defined can be conveniently prepared byreaction of a compound of formula (XXI)

wherein R₁ is R′ and X is as defined above, with a compound of formula(XX) as defined above. The reaction can be carried out in the presenceof a base such as triethylamine, N,N-diisopropylethylamine, pyridine orsodium hydride in a suitable solvent such as toluene, diethyl ether,tetrahydrofuran, acetonitrile, dimethyl sulfoxide orN,N-dimethylformamide, at a temperature ranging from about 10° C. toreflux and for a time varying from about 30 minutes to about 96 hoursand, subsequently, by treating the obtained product with zinc powder inacetic acid or magnesium powder in methanol, at a temperature rangingfrom about 10° C. to 60° C. and for a time varying from about 30 minutesto about 96 hours.

Finally, from all of the above, it is also clear to the person skilledin the art that the compounds of formula (I) of the invention can bealso prepared by performing the reactions described above in acombinatorial fashion.

As an example, the compounds of formula (VII) supported onto resinparticles, prepared as above described, may be reacted with a variety ofcompounds of formula from (IX) to (XVI) so as to obtain thousands ofdifferent compounds of formula (VIII), according to combinatorialchemistry methods. These latter derivatives, in their turn, may be thenconveniently converted into the derivatives of formula (I) of theinvention.

It is therefore a further object of the invention a combinatorialchemical library comprising a plurality of members of formula (VIII)

wherein

-   X is NR′, O, S, SO or SO₂;-   each of R and R₁, being the same or different, is independently a    hydrogen atom or an optionally substituted group selected from R′,    —COR′, —COOR′, —CONHR′, —CONR′R″, —SO₂R′, —SO₂NHR′ or —SO₂NR′R″;    wherein each of R′ and R″, being the same or different, is    independently selected from hydrogen or an optionally further    substituted straight or branched C₁-C₆ alkyl, heterocyclyl, aryl or    aryl-C₁-C₆ alkyl group;-   R₂ is an optionally substituted group selected from R′, —CH₂OR′ and    OR′, wherein R′ is as above defined; and the pharmaceutically    acceptable salts thereof.

Preferably, the above resin is a polystrenic resin, in particular amethylisocyanate polystyrene resin.

The said resin may be then cleaved by working according to conventionalmethods so as to give rise to a plurality of compounds of formula (I).

Therefore, it is a further object of the invention a chemical library ofcompounds comprising two or more derivatives of formula (I)

wherein

-   X is NR′, O, S, SO or SO₂;-   each of R and R₁, being the same or different, is independently a    hydrogen atom or an optionally substituted group selected from R′,    —COR′, —COOR′, —CONHR′, —CONR′R″, —SO₂R′, —SO₂NHR′ or —SO₂NR′R″;    wherein each of R′ and R″, being the same or different, is    independently selected from hydrogen or an optionally further    substituted straight or branched C₁-C₆ alkyl, heterocyclyl aryl or    aryl-C₁-C₆ alkyl group;-   R₂ is an optionally substituted group selected from R′, —CH₂OR′ and    OR′, wherein R′ is as above defined; and the pharmaceutically    acceptable salts thereof.

From all of the above, it is clear to the skilled person that once alibrary of compounds of formula (I) is thus prepared, for instanceconsisting of several hundreds of members, the said library can be veryadvantageously used for screening towards given kinases, as formerlyreported.

See, for a general reference to libraries of compounds and uses thereofas tools for screening biological activities, J. Med. Chem. 1999, 42,2373-2382; and Bioorg. Med. Chem. Lett. 10 (2000), 223-226.

Pharmacology

The compounds of formula (I) are active as protein kinase inhibitors andare therefore useful, for instance, to restrict the unregulatedproliferation of tumor cells.

In particular they are active as cyclin dependent kinase (cdk)inhibitors and also as inhibitors of other protein kinases such as, forinstance, protein kinase C in different isoforms, Met, PAK-4, PAK-5,ZC-1, STLK-2, DDR-2, Aurora 1, Aurora 2, Bub-1, PLK, Chk1, Chk2, HER2,raf1, MEK1, MAPK, EGF-R, PDGF-R, FGF-R, IGF-R, VEGF-R, PI3K, wee1kinase, Src, Abl, Akt, ILK, MK-2, IKK-2, Cdc7, Nek, and thus to beeffective in the treatment of diseases associated with other proteinkinases.

In therapy, they may be used in the treatment of various tumors, such asthose formerly reported, as well as in the treatment of other cellproliferative disorders such as psoriasis, vascular smooth cellproliferation associated with atherosclerosis and post-surgical stenosisand restenosis and in the treatment of Alzheimer's disease.

The inhibiting activity of putative cdk/cyclin inhibitors and thepotency of selected compounds is determined through a method of assaybased on the use of the SPA technology (Amersham Pharmacia Biotech).

The assay consists of the transfer of radioactivity labelled phosphatemoiety by the kinase to a biotinylated substrate. The resulting33P-labelled biotinylated product is allowed to bind tostreptavidin-coated SPA beads (biotin capacity 130 pmol/mg), and lightemitted was measured in a scintillation counter.

Inhibition Assay of cdk2/Cyclin A Activity

Kinase reaction: 4 μM in house biotinylated histone H1 (Sigma # H-5505)substrate, 10 μM ATP (0.1 microCi P³³γ-ATP), 1.1 nM Cyclin A/CDK2complex, inhibitor in a final volume of 30 μl buffer (TRIS HCl 10 mM pH7.5, MgCl₂ 10 mM, DTT 7.5 mM+0.2 mg/ml BSA) were added to each well of a96 U bottom. After incubation for 60 min at room temperature, thereaction was stopped by addition of 100 μl PBS buffer containing 32 mMEDTA, 500 μM cold ATP, 0.1% Triton X100 and 10 mg/ml streptavidin coatedSPA beads. After 20 min incubation, 110 μL of suspension were withdrawnand transferred into 96-well-OPTIPLATEs containing 100 μl of 5M CsCl.After 4 hours, the plates were read for 2 min in a Packard TOP-Countradioactivity reader.

IC50 determination: inhibitors were tested at different concentrationsranging from 0.0015 to 10 μM. Experimental data were analyzed by thecomputer program GraphPad Prizm using the four parameter logisticequation:y=bottom+(top-bottom)/(1+10^((log IC50-x)*slope))where x is the logarithm of the inhibitor concentration, y is theresponse; y starts at bottom and goes to top with a sigmoid shape.Ki Calculation:

Experimental method: Reaction was carried out in buffer (10 mM Tris, pH7.5, 10 mM MgCl₂, 0.2 mg/ml BSA, 7.5 mM DTT) containing 3.7 nM enzyme,histone and ATP (constant ratio of cold/labeled ATP 1/3000). Reactionwas stopped with EDTA and the substrate captured on phosphomembrane(Multiscreen 96 well plates from Millipore). After extensive washing,the multiscreen plates were read on a top counter. Control (time zero)for each ATP and histone concentrations was measured.

Experimental design: Reaction velocities are measured at four ATP,substrate (histone) and inhibitor concentrations. An 80-pointconcentration matrix was designed around the respective ATP andsubstrate Km values, and the inhibitor IC50 values (0.3, 1, 3, 9 foldthe Km or IC50 values). A preliminary time course experiment in theabsence of inhibitor and at the different ATP and substrateconcentrations allows the selection of a single endpoint time (10 min)in the linear range of the reaction for the Ki determination experiment.

Kinetic parameter estimates: Kinetic parameters were estimated bysimultaneous nonlinear least-square regression using [Eq. 1](competitive inhibitor respect to ATP, random mechanism) using thecomplete data set (80 points):

$\begin{matrix}{v = \frac{{Vm} \cdot A \cdot B}{\begin{matrix}{{\alpha \cdot {Ka} \cdot {Kb} \cdot {+ \alpha} \cdot {Ka} \cdot B} + {a \cdot}} \\{{{Kb} \cdot A} + {A \cdot B} + {\alpha \cdot \;\frac{Ka}{Ki} \cdot I \cdot \left( {{Kb} + \frac{B}{\beta}} \right)}}\end{matrix}}} & \left\lbrack {{Eq}.\mspace{14mu} 1} \right\rbrack\end{matrix}$where A=[ATP], B=[Substrate], I=[inhibitor], Vm=maximum velocity, Ka,Kb, Ki the dissociation constants of ATP, substrate and inhibitorrespectively. α and β the cooperativity factor between substrate and ATPbinding and substrate and inhibitor binding respectively.

In addition the selected compounds are characterized on a panel ofser/thre kinases strictly related to cell cycle (cdk2/cyclin E,cdk1/cyclin B1, cdk5/p25, cdk4/cyclin D1), and also for specificity onMAPK, PKA, EGFR, IGF1-R, Aurora-2 and Cdc 7.

Inhibition Assay of cdk2/Cyklin E Activity

Kinase reaction: 10 μM in house biotinylated histone H1 (Sigma # H-5505)substrate, 30 μM ATP (0.3 microCi P³³γ-ATP), 4 ng GST-Cyclin E/CDK2complex, inhibitor in a final volume of 30 μl buffer (TRIS HCl 10 mM pH7.5, MgCl₂ 10 mM, DTT 7.5 mM+0.2 mg/ml BSA) were added to each well of a96 U bottom. After incubation for 60 min at room temperature, thereaction was stopped by addition of 100 μl PBS buffer containing 32 mMEDTA, 500 μM cold ATP, 0.1% Triton X100 and 10 mg/ml streptavidin coatedSPA beads. After 20 min incubation, 110 μL of suspension were withdrawnand transferred into 96-well OPTIPLATEs containing 100 μl of 5M CsCl.After 4 hours, the plates were read for 2 min in a Packard TOP-Countradioactivity reader.

IC50 determination: see above

Inhibition Assay of cdk1/Cyclin B1 Activity

Kinase reaction: 4 μM in house biotinylated histone H1 (Sigma # H-5505)substrate, 20 μM ATP (0.2 microCi P³³γ-ATP), 3 ng Cyclin B/CDK1 complex,inhibitor in a final volume of 30 μl buffer (TRIS HCl 10 mM pH 7.5,MgCl₂ 10 mM, DTT 7.5 mM+0.2 mg/ml BSA) were added to each well of a 96 Ubottom. After 20 min at r.t. incubation, reaction was stopped by 100 μlPBS+32 mM EDTA+0.1% Triton X-100+500 μM ATP, containing 1 mg SPA beads.Then a volume of 110 μl is transferred to Optiplate. After 20 min.incubation for substrate capture, 100 μl 5M CsCl were added to allowstatification of beads to the top of the Optiplate and let stand 4 hoursbefore radioactivity counting in the Top-Count instrument.

IC50 determination: see above

Inhibition Assay of cdk5/p25 Activity

The inhibition assay of cdk5/p25 activity is performed according to thefollowing protocol.

Kinase reaction: 10 μM biotinylated histone H1 (Sigma # H-5505)substrate, 30 μM ATP (0.3 microCi P³³γ-ATP), 15 ng CDK5/p25 complex,inhibitor in a final volume of 30 μl buffer (TRIS HCl 10 mM pH 7.5,MgCl₂ 10 mM, DTT 7.5 mM+0.2 mg/ml BSA) were added to each well of a 96 Ubottom. After incubation for 35 min at room temperature, the reactionwas stopped by addition of 100 μl PBS buffer containing 32 mM EDTA, 500μM cold ATP, 0.1% Triton X100 and 10 mg/ml streptavidin coated SPAbeads. After 20 min incubation, 110 μL of suspension were withdrawn andtransferred into 96-well OPTIPLATEs containing 100 μl of 5M CsCl. After4 hours, the plates were read for 2 min in a Packard TOP-Countradioactivity reader.

IC50 determination: see above

Inhibition Assay of cdk4/Cyclin D1 Activity

Kinase reaction: 0.4 uM μM mouse GST-Rb (769-921) (# sc-4112 from SantaCruz) substrate, 10 μM ATP (0.5 μCi P³³γ-ATP), 100 ng of baculovirusexpressed GST-cdk4/GST-Cyclin D1, suitable concentrations of inhibitorin a final volume of 50 μl buffer (TRIS HCl 10 mM pH 7.5, MgCl₂ 10 mM,7.5 mM DTT+ 0.2 mg/ml BSA) were added to each well of a 96 U bottom wellplate. After 40 min at 37° C. incubation, reaction was stopped by 20 μlEDTA 120 mM.

Capture: 60 μl were transferred from each well to MultiScreen plate, toallow substrate binding to phosphocellulose filter. Plates were thenwashed 3 times with 150 μl/well PBS Ca⁺⁺/Mg⁺⁺ free and filtered byMultiScreen filtration system.

Detection: filters were allowed to dry at 37° C., then 100 μl/wellscintillant were added and ³³P labeled Rb fragment was detected byradioactivity counting in the Top-Count instrument.

IC50 determination: see above

Inhibition Assay of MAPK Activity

Kinase reaction: 10 μM in house biotinylated MBP (Sigma # M-1891)substrate, 15 μM ATP (0.15 microCi P³³γ-ATP), 30 ng GST-MAPK UpstateBiothecnology # 14-173), inhibitor in a final volume of 30 μl buffer(TRIS HCl 10 mM pH 7.5, MgCl₂ 10 mM, DTT 7.5 mM+0.2 mg/ml BSA) wereadded to each well of a 96 U bottom. After incubation for 35 min at roomtemperature, the reaction was stopped by addition of 100 μl PBS buffercontaining 32 mM EDTA, 500 μM cold ATP, 0.1% Triton X100 and 10 mg/mlstreptavidin coated SPA beads. After 20 min incubation, 110 μL ofsuspension were withdrawn and transferred into 96-well OPTIPLATEscontaining 100 μl of 5M CsCl. After 4 hours, the plates were read for 2min in a Packard TOP-Count radioactivity reader.

IC50 determination: see above

Inhibition Assay of PKA Activity

Kinase reaction: 10 μM in house biotinylated histone H1 (Sigma # H-5505)substrate, 10 μM ATP (0.2 microM P³³γ-ATP), 0.45 U PKA (Sigma # 2645),inhibitor in a final volume of 30 μl buffer (TRIS HCl 10 mM pH 7.5,MgCl₂ 10 mM, DTT 7.5 mM+0.2 mg/ml BSA) were added to each well of a 96 Ubottom. After incubation for 90 min at room temperature, the reactionwas stopped by addition of 100 μl PBS buffer containing 32 mM EDTA, 500μM cold ATP, 0.1% Triton X100 and 10 mg/ml streptavidin coated SPAbeads. After 20 min incubation, 110 μL of suspension were withdrawn andtransferred into 96-well OPTIPLATEs containing 100 μl of 5M CsCl. After4 hours, the plates were read for 2 min in a Packard TOP-Countradioactivity reader.

IC50 determination: see above

Inhibition Assay of EGFR Activity

Kinase reaction: 10 μM in house biotinylated MBP (Sigma # M-1891)substrate, 2 μM ATP (0.04 microCi P³³γ-ATP), 36 ng insect cell expressedGST-EGFR, inhibitor in a final volume of 30 μl buffer (Hepes 50 mM pH7.5, MgCl₂ 3 mM, MnCl₂ 3 mM, DTT 1 mM, NaVO₃ 3 μM, +0.2 mg/ml BSA) wereadded to each well of a 96 U bottom. After incubation for 20 min at roomtemperature, the reaction was stopped by addition of 100 μl PBS buffercontaining 32 mM EDTA, 500 μM cold ATP, 0.1% Triton X100 and 10 mg/mlstreptavidin coated SPA beads. After 20 min incubation, 110 μL ofsuspension were withdrawn and transferred into 96-well OPTIPLATEscontaining 100 μl of 5M CsCl. After 4 hours, the plates were read for 2min in a Packard TOP-Count radioactivity reader.

IC50 determination: see above

Inhibition Assay of XGF1-R Activity

The inhibition assay of IGF1-R activity is performed according to thefollowing protocol.

Enzyme activation: IGF1-R must be activated by auto-phosphorylationbefore starting the experiment. Just prior to the assay, a concentratedenzyme solution (694 nM) is incubated for half a hour at 28° C. in thepresence of 100 μM ATP and then brought to the working dilution in theindicated buffer.

Kinase reaction: 10 μM biotinylated IRS1 peptide (PRIMM) substrate, 0-20μM inhibitor, 6 μM ATP, 1 microCi ³³P-ATP, and 6 nM GST-IGF1-R(pre-incubated for 30 min at room temperature with cold 60 μM cold ATP)in a final volume of 30 μl buffer (50 mM HEPES pH 7.9, 3 mM MnCl₂, 1 mMDTT, 3 μM NaVO₃) were added to each well of a 96 U bottom well plate.After incubation for 35 min at room temperature, the reaction wasstopped by addition of 100 μl PBS buffer containing 32 mM EDTA, 500 μMcold ATP, 0.1% Triton X100 and 10 mg/ml streptavidin coated SPA beads.After 20 min incubation, 110 μL of suspension were withdrawn andtransferred into 96-well OPTIPLATEs containing 100 μl of 5M CsCl. After4 hours, the plates were read for 2 min in a Packard TOP-Countradioactivity reader.

Inhibition Assay of Aurora-2 Activity

Kinase reaction: 8 μM biotinylated peptide (4 repeats of LRRWSLG), 10 μMATP (0.5 uCi P³³γ-ATP), 7.5 ng Aurora 2, inhibitor in a final volume of30 μl buffer (HEPES 50 mM pH 7.0, MgCl₂ 10 mM, 1 mM DTT, 0.2 mg/ml BSA,3 μM orthovanadate) were added to each well of a 96 U bottom well plate.After 60 minutes at room temperature incubation, reaction was stoppedand biotinylated peptide captured by adding 100 μl of bead suspension.

Stratification: 100 μl of CsCl2 5 M were added to each well and letstand 4 hour before radioactivity was counted in the Top-Countinstrument.

IC50 determination: see above

Inhibition Assay of Cdc7/dbf4 Activity

The inhibition assay of Cdc7/dbf4 activity is performed according to thefollowing protocol.

The Biotin-MCM2 substrate is trans-phosphorylated by the Cdc7/Dbf4complex in the presence of ATP traced with γ³³-ATP. The phosphorylatedBiotin-MCM2 substrate is then captured by Streptavidin-coated SPA beadsand the extent of phosphorylation evaluated by β counting.

The inhibition assay of Cdc7/dbf4 activity was performed in 96 wellsplate according to the following protocol.

To each well of the plate were added:

-   -   10 μl substrate (biotinylated MCM2, 6 μM final concentration)    -   10 μl enzyme (Cdc7/Dbf4, 17.9 nM final concentration)    -   10 μl test compound (12 increasing concentrations in the nM to        μM range to generate a dose-response curve)    -   10 μl of a mixture of cold ATP (2 μM final concentration) and        radioactive ATP (1/5000 molar ratio with cold ATP) was then used        to start the reaction which was allowed to take place at 37° C.

Substrate, enzyme and ATP were diluted in 50 mM HEPES pH 7.9 containing15 mM MgCl₂, 2 mM DTT, 3 μM NaVO₃, 2 mM glycerophosphate and 0.2 mg/mlBSA. The solvent for test compounds also contained 10% DMSO.

After incubation for 60 minutes, the reaction was stopped by adding toeach well 100 μl of PBS pH 7.4 containing 50 mM EDTA, 1 mM cold ATP,0.1% Triton X100 and 10 mg/ml streptavidin coated SPA beads.

After 20 min incubation, 110 μL of suspension were withdrawn andtransferred into 96-well OPTIPLATEs containing 100 μl of 5M CsCl. After4 hours, the plates were read for 2 min in a Packard TOP-Countradioactivity reader.

IC50 determination: see above.

In view of the above biological activities the compounds of theinvention can be useful in therapy, for instance, to restrict theunregulated proliferation of tumor cells. More specifically, thebicyclo-pyranzoles of the invention can be useful in the treatment of avariety of cancers including, but not limited to carcinoma of severalorgans, tissues and glands such as bladder, breast, colon, kidney,liver, lung, including small cell lung cancers esophagus, gall-bladder,ovary, pancreas, stomach, cervix, thyroid, prostate, and skin, includingsquamous cell carcinoma; hematopoietic tumors of lymphoid lineage,including leukemia, acute lymphocitic leukemia, acute lymphoblasticleukemia, B-cell lymphoma, T-cell-lymphoma, Hodgkin's lymphoma,non-Hodgkin's lymphoma, hairy cell lymphoma and Burkett's lymphoma;hematopoietic tumors of myeloid lineage, including acute and chronicmyelogenous leukemias, myelodysplastic syndrome and promyelocyticleukemia; tumors of mesenchymal origin, including fibrosarcoma andrhabdomyosarcoma; tumors of the central and peripheral nervous system,including astrocytoma, neuroblastoma, glioma and schwannomas; othertumors, including melanoma, seminoma, teratocarcinoma, osteosarcoma,xeroderma pigmentosum, keratoxanthomas, thyroid follicular cancer andKaposi's sarcoma. Due to the key role of PKs in the regulation ofcellular proliferation, the bicyclo-pyrazoles of the invention can alsobe useful in the treatment of a variety of cell proliferative disorderssuch as, for instance, benign prostate hyperplasia, familialadenomatosis, polyposis, neuro-fibromatosis, psoriasis, vascular smoothcell proliferation associated with atherosclerosis, pulmonary fibrosis,arthritis glomerulonephritis and post-surgical stenosis and restenosis.The compounds of the invention can be useful in the treatment ofAlzheimer's disease, as suggested by the fact that cdk5 is involved inthe phosphorylation of tau protein (J. Biochem., 117, 741-749, 1995).The compounds of the invention, as modulators of apoptosis, may also beuseful in the treatment of cancer, viral infections, prevention of AIDSdevelopment in HIV-infected individuals, autoimmune diseases andneurodegenerative disorders. The compounds of the invention may beuseful in inhibiting tumor angiogenesis and metastasis.

The compounds of the invention are suitable for administration to amammal, e.g. to humans, by the usual routes. The dosage level depends asusual upon the age, weight, conditions of the patient and theadministration route.

For example, a suitable dosage adopted for oral administration of thecompounds of the invention, for instance,N-benzyl-3-{[(4-ethylpiperazin-1-yl)benzoyl]amino}-1H-thieno[3,2-c]pyrazole-5-carboxamide,may range from about 5 to about 500 mg pro dose, from 1 to 5 timesdaily.

The compounds of the invention can be administered in a variety ofdosage forms, e.g. orally, in the form of tablets, capsules, sugar orfilm coated tablets, liquid solutions or suspensions; rectally in theform of suppositories; parenterally, e.g. intramuscularly, byintravenous and/or intrathecal and/or intraspinal injection or infusion;or by transdermal administration.

In addition, the compounds of the invention can be administered eitheras single agents or, alternatively, in a combination therapy methodcomprising additional anticancer treatments such as radiation therapy orchemotherapy regimen in combination with cytostatic or cytotoxic agents,antibiotic-type agents, alkylating agents, antimetabolite agents,hormonal agents, immunological agents, interferon-type agents,cyclooxygenase inhibitors (e.g. COX-2 inhibitors, in particularcelecoxib, rofecoxib, parecoxib and valdecoxib), metallomatrixproteaseinhibitors, telomerase inhibitors, tyrosine kinase inhibitors,anti-growth factor receptor agents, anti-HER agents, anti-EGFR agents,anti-angiogenesis agents, farnesyl transferase inhibitors, ras-rafsignal transduction pathway inhibitors, cell cycle inhibitors, othercdks inhibitors, tubulin binding agents, topoisomerase I inhibitors,topoisomerase II inhibitors, and the like.

If formulated as a fixed dose, such combination products employ thecompounds of his invention within the dosage range described above andthe other pharmaceutically active agent within the approved dosagerange.

Compounds of formula (I) may be used sequentially with known anticanceragents when a combination formulation is inappropriate.

The invention, therefore, also provides a method for treating a mammal,including humans, suffering from a disease caused by and/or associatedwith an altered (disregulated) protein kinase activity, comprisingadministering to said mammal in need thereof a therapeutically effectiveamount of a bicyclo-pyrazole compound of formula (I), or apharmaceutically acceptable salt thereof, while undergoing simultaneous,separate or sequential anticancer treatments.

A further object of the invention is the use of a compound of formula(I), or a pharmaceutically acceptable salt thereof, in the manufactureof a medicament for treating a disease caused by and/or associated withan altered protein kinase activity, in a patient undergoing asimultaneous, separate or sequential anticancer treatments.

The present invention also includes pharmaceutical compositionscomprising a compound of formula (I) or a pharmaceutically acceptablesalt thereof in association with a pharmaceutically acceptableexcipient, which can be a carrier or a diluent.

The pharmaceutical compositions containing the compounds of theinvention are usually prepared following conventional methods and areadministered in a pharmaceutically suitable form.

For example, the solid oral forms may contain, together with the activecompound, diluents, e.g. lactose, dextrose, saccharose, sucrose,cellulose, corn starch or potato starch; lubricants, e.g. silica, talc,stearic, magnesium or calcium stearate, and/or polyethylene glycols;binding agents, e.g. starches, arabic gum, gelatine, methylcellulose,carboxymethylcellulose or polyvinyl pyrrolidone; disaggregating agents,e.g. a starch, alginic, alginates or sodium starch glycolate;effervescing mixtures; dyestuffs; sweeteners; wetting agents such aslecithin, polysorbates, laurylsulphates; and, in, general, non-toxic andpharmacologically inactive substances used in pharmaceuticalformulations. Said pharmaceutical preparations may be manufactured inknown manner, for example, by means of mixing, granulating, tabletting,sugar-coating, or film-coating processes.

The liquid dispersions for oral administration may be e.g. syrups,emulsions and suspensions.

The syrups may contain as carrier, for example, saccharose or saccharosewith glycerine and/or mannitol and/or sorbitol.

The suspensions and the emulsions may contain as carrier, for example, anatural gum, agar, sodium alginate, pectin, methylcellulose,carboxymethylcellulose, or polyvinyl alcohol.

The suspension or solutions for intramuscular injections may contain,together with the active compound, a pharmaceutically acceptablecarrier, e.g. sterile water, olive oil, ethyl oleate, glycols, e.g.propylene glycol, and, if desired, a suitable amount of lidocainehydrochloride. The solutions for intravenous injections or infusions maycontain as carrier, for example, sterile water or preferably they may bein the form of sterile, aqueous, isotonic saline solutions or they maycontain as a carrier propylene glycol.

The suppositories may contain together with the active compound apharmaceutically acceptable carrier, e.g. cocoa butter, polyethyleneglycol, a polyoxyethylene sorbitan fatty ester surfactant or lecithin.

The following examples are herewith intended to illustrate, withoutposing any limitation to, the present invention.

SYNTHETIC EXAMPLES

The following HPLC methods were used in the analysis of the compounds asspecified in the synthetic examples set forth below. As used herein, theterm “Rt” refers to the retention time for the compound using the HPLCmethod specified.

Method A

HPLC/MS was performed on a Waters X Terra RP 18 (4.6×50 mm, 3.5 μm)column using a Waters 2790 HPLC system equipped with a 996 Waters PDAdetector and a Micromass mod. ZQ single quadrupole mass spectrometer,equipped with an electrospray (ESI) ion source. Mobile phase A wasammonium acetate 5 mM buffer (pH 5.5 with acetic acid/acetonitrile95:5), and Mobile phase B was H2O/acetonitrile (5:95). Gradient from 10to 90% B in 8 minutes, hold 90% B 2 min. UV detection at 220 nm and 254nm. Flow rate 1 ml/min. Injection volume 10 μl. Full scan, mass rangefrom 100 to 800 amu. Capillary voltage was 2.5 KV; Source temp was 120°C.; Cone was 10 V. Retention Times (HPLC r.t.) are given in minutes at220 nm or 254 nm. Mass are given as m/z ratio.

Method B

HPLC/MS was performed on a Hypersil C18 BDS (2×50 mm, 5 μm) column usinga Hewlett Packard 1312A HPLC system equipped with a Polymer Labs PL1000Evaporative Light Scattering detector and a Micromass ZMD massspectrometer, equipped with an electrospray (ESI) ion source. Mobilephase A was aqueous solution of trifluoroacetic acid (0.1% v/v), andMobile phase B was acetonitrile solution of trifluoroacetic acid (0.1%v/v). Gradient from 0 to 95% B in 1.8 minutes, hold 95% B for 0.3 min.Flow rate 1 ml/min. Injection volume 3 μl. Full scan, mass range from150 to 800 amu. Source temp was 140° C.; Cone was 25 V. Retention Times(HPLC r.t.) are given in minutes. Mass are given as m/z ratio.

Example 1 4-cyano-5-(methylthio)thiophene-2-carboxylic acid

Aqueous sodium hydroxide (20% w/w solution, 9 mL) was added to asolution of ethyl 4-cyano-5-(methylthio)thiophene-2-carboxylate (10 g,44 mmol) in 1,4-dioxane (100 mL) at 5° C.

After stirring for 4 hours at room temperature, water (500 mL) was addedto the reaction mixture and the pH was adjusted to pH about 2.5 byadding 2N aqueous hydrochloric acid. A white solid was separated byfiltration, washed with water, and dried under vacuum to give 8.5 g ofthe title compound.

Chromatographic method A, Rf 2.4; [M+H]⁺ 200.

Example 2 tert-butyl 4-cyano-5-(methylthio)thiophene-2-carboxylate

A mixture of 4-cyano-5-(methylthio)thiophene-2-carboxylic acid (2.0 g,10 mmol), benzyltrimethylammonium chloride (2.25 g, 10 mmol), tertbutylbromide (54 mL, 480 mmol), and anhydrous potassium carbonate (36 g, 260mmol) in dimethylacetamide (100 mL) was stirred at 60° C. for 6 hours.

After cooling the mixture was diluted with ethyl acetate (400 mL) andwashed with water. Organic layer was dried and evaporated under reducedpressure to give a residue which was purified by chromatography (eluentethyl acetate/n-hexane 3:1) yielding 1.5 g of the title compound.

Chromatographic method A, Rf 7.4; [M+H]⁺ 256.

Example 3 tert-butyl 4-cyano-5-(methylsulfonyl)thiophene-2-carboxylate

A mixture of tert-butyl 4-cyano-5-(methylthio)thiophene-2-carboxylate(1.4 g, 5.5 mmol) and oxone (14.4 g, 21.5 mmol) in dimethylformamide(100 mL) was stirred at room temperature for 16 hours. The reactionmixture was then poured into ice/water (400 mL) and extracted with ethylacetate. Organic layer was washed with water, dried over anhydroussodium sulfate and evaporated to give 1.5 g of the title compound.

Chromatographic method A, Rf 6.2; [M+H]⁺ 288.

Example 4 tert-butyl 4-cyano-5-hydrazinothiophene-2-carboxylate

A mixture of tert-butyl 4-cyano-5-methylsulfonyl)thiophene-2-carboxylate (2.0 g, 7.0 mmol) and hydrazine hydrate (1.7 mL)in methyl alcohol (30 mL) was stirred at 60° C. for 2 hours. Thereaction mixture was diluted with ethyl acetate (100 mL) and washed withwater. Organic layer was separated, dried over anhydrous sodium sulfate,and evaporated. Purification by chromatography (n-hexane/ethyl acetate3:2) gave 1 g of the title compound.

Chromatographic method A, Rf 5.6; [M+H]⁺ 240.

Example 5 tert-butyl 3-amino-1H-thieno[2,3-c]pyrazole-5-carboxylate

A mixture of tert-butyl 4-cyano-5-hydrazinothiophene-2-carboxylate (1.0g, 4.2 mmol) and hydrochloric acid (0.7 mL of 37% solution) in methylalcohol (15 mL) was stirred at room temperature for 14 hours. Thereaction mixture was diluted with ethyl acetate (50 mL) and washed withan aqueous solution of sodium bicarbonate. Organic layer was separated,dried over anhydrous sodium sulfate, and evaporated to give 0.9 g of thetitle compound.

Chromatographic method A, Rf 4.5; [M+H]⁺ 240.

Example 6

By working as described in the previous examples and by starting fromany suitable starting material, according to processes object of theinvention, the following compounds of formula (I) can be obtained:

-   -   1.        N-benzyl-3-[(4-fluorobenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   2.        N-ethyl-3-{[4-(4-ethylpiperazin-1-yl)benzoyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   3.        3-{[4-(4-ethylpiperazin-1-yl)benzoyl]amino}-N-isopropyl-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   4.        N-benzyl-3-{[4-(4-ethylpiperazin-1-yl)benzoyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   5.        N-{5-[(4-methylpiperazin-1-yl)carbonyl]-1H-thieno[2,3-c]pyrazol-3-yl}benzamide;    -   6. N-[5-morpholin        ylcarbonyl)-1H-thieno[2,3-c]pyrazol-3-yl]benzamide;    -   7.        3-(benzoylamino)-N-2-morpholin-4-ylethyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   8.        3-(benzoylamino)-N-phenyl-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   9.        3-(benzoylamino)-N-(2-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   10.        3-(benzoylamino)-N-(4-cyanophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   11.        3-(benzoylamino)-N-(4-fluorophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   12.        3-(benzoylamino)-N-(4-chlorophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   13.        3-(benzoylamino)-N-(4-methoxyphenyl)1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   14.        3-(benzoylamino)-N-(4-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   15.        3-(benzoylamino)-N-tert-butyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   16.        3-(benzoylamino)-N-isopropyl-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   17.        3-benzoylamino)-N-ethyl-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   18.        2-fluoro-N-{5-[(4-methylpiperazin-1-yl)carbonyl]-1H-thieno[2,3-c]pyrazol-3-yl}benzamide;    -   19.        2-fluoro-N-[5-(morpholin-4-ylcarbonyl)-1H-thieno[2,3-c]pyrazol-3-yl]benzamide;    -   20.        3-[(2-fluorobenzoyl)amino]-N-(2-morpholin-4-ylethyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   21.        3-[(2-fluorobenzoyl)amino]-N-phenyl-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   22.        3-[(2-fluorobenzoyl)amino]-N-(2-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   23.        N-(4-cyanophenyl)-3-[(2-fluorobenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   24.        3-[(2-fluorobenzoyl)amino]-N-(4-fluorophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   25.        N-(4-chlorophenyl)-3-[(2-fluorobenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   26.        3-[(2-fluorobenzoyl)amino]-N-(4-methoxyphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   27.        3-[(2-fluorobenzoyl)amino]-N-(4-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   28.        N-tert-butyl)-3-[(2-fluorobenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   29.        3-[(2-fluorobenzoyl)amino]-N-isopropyl-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   30. N        ethyl-3-[(2-fluorobenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   31.        2-chloro-N-{5-[(4-methylpiperazin-1-yl)carbonyl]-1H-thieno[2,3-c]pyrazol-3-yl}benzamide;    -   32.        2-chloro-N-[5-(morpholin-4-ylcarbonyl)-1H-thieno[2,3-c]pyrazol-3-yl]benzamide;    -   33.        3-[(2-chlorobenzoyl)amino]-N-(2-morpholinylethyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   34.        3-[(2-chlorobenzoyl)amino]-N-phenyl-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   35.        3-[(2-chlorobenzoyl)amino]-N-(2-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   36.        3-[(2-chlorobenzoyl)amino]-N-(4-cyanophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   37.        3-[(2-chlorobenzoyl)amino]-N-(4-fluorophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   38.        3-[(2-chlorobenzoyl)amino]-N-(4-chlorophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   39.        3-[(2-chlorobenzoyl)amino]-N-(4-methoxyphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   40.        3-[(2-chlorobenzoyl)amino]-N-(4-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   41.        N-(tert-butyl)-3-[(2-chlorobenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   42.        3-[(2-chlorobenzoyl)amino]-N-isopropyl-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   43.        3-[(2-chlorobenzoyl)amino]-N-ethyl-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   44.        2-methoxy-N-{5-[(4-methylpiperazin-1-yl)carbonyl]-1H-thieno[2,3-c]pyrazol-3-yl}benzamide;    -   45.        2-methoxy-N-[5-(morpholin-4-ylcarbonyl)-1H-thieno[2,3]pyrazol-3-yl]benzamide;    -   46.        3-[(2-methoxybenzoyl)amino]-N-(2-morpholin-4-ylethyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   47.        3-[(2-methoxybenzoyl)amino]-N-phenyl-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   48.        3-[(2-methoxybenzoyl)amino]-N-(2-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   49.        N-(4-cyanophenyl)-3-[(2-methoxybenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   50.        N-(4-fluorophenyl)-3-[(2-methoxybenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   51.        N-(4-chlorophenyl)-3-[(2-methoxybenzoyl)amino]-1H-thieno[2,3c]pyrazole-5-carboxamide;    -   52.        3-[(2-methoxybenzoyl)amino]-N-(4-methoxyphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   53.        3-[(2-methoxybenzoyl)amino]-N-(4-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   54.        N-(tert-butyl)-3-[(2-methoxybenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   55.        N-isopropyl-3-[(2-methoxybenzoyl)amino]-1H-thieno[2,]pyrazole-5-carboxamide;    -   56.        N-ethyl-3-[(2-methoxybenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   57.        2-methyl-N-{5-[(4-methylpiperazin-1-yl)carbonyl]-1H-thieno[2,3-c]pyrazol-3-yl}benzamide;    -   58.        2-methyl-N-[5-(morpholin-4-ylcarbonyl)-1H-thieno[2,3-c]pyrazol-3-yl]benzamide;    -   59.        3-[(2-methylbenzoyl)amino]-N-(2-morpholin-4-ylethyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   60.        3-[(2-methylbenzoyl)amino]-N-phenyl-1H-thieno[2,3-c]pyrazole-5        carboxamide;    -   61.        3-[(2-methylbenzoyl)amino]-N-(2-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   62.        N-(4-cyanophenyl)-3-[(2-methylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   63.        N-(4-fluorophenyl)-3-[(2-methylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   64.        N-(4-chlorophenyl)-3-[(2-methylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   65.        N-(4-methoxyphenyl)-3-[(2-methylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   66.        3-[(2-methylbenzoyl)amino]-N-(4-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   67.        N-(tert-butyl)-3-[(2-methylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   68.        N-isopropyl-3-[(2-methylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   69.        N-ethyl-3-[(2-methylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   70.        3-methyl-N-{5-[(4-methylpiperazin-1-yl)carbonyl]-1H-thieno[2,3-c]pyrazol-3-yl}benzamide;    -   71.        3-methyl-N-[5-(morpholin-4-ylcarbonyl)-1H-thieno[2,3-c]pyrazol-3-yl]benzamide;    -   72.        3-[(3-methylbenzoyl)amino]-N-(2-morpholin-4-ylethyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   73.        3-[(3-methylbenzoyl)amino]-N-phenyl-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   74.        3-[3-methylbenzoyl)amino]-N-(2-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   75.        N-(4-cyanophenyl)-3-[(3-methylbenzyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   76.        N-(4-fluorophenyl)-3-[(3-methylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   77.        N-(4-chlorophenyl)-3-[(3-methylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   78.        N-(4-methoxyphenyl)-3-[(3-methylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   79.        3-[(3-methylbenzoyl)amino]-N-(4-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   80.        N-(tert-butyl)-3-[(3-methylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   81.        N-isopropyl-3-[(3-methylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   82.    N-ethyl-3-[(3-methylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   83.        4-fluoro-N-{5-[(4-methylpiperazin-1-yl)carbonyl]-1H-thieno[2,3-c]pyrazol-3-yl}benzamide;    -   84.        4-fluoro-N-[5-morpholin-4-ylcarbonyl)-1H-thieno[2,3-c]pyrazol-3-yl]benzamide;    -   85.        3-[(4-fluorobenzoyl)amino]-N-(2-morpholin-4-ylethyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   86.        3-[(4-fluorobenzoyl)amino]-N-phenyl-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   87.        3-[(4-fluorobenzoyl)amino]-N-(2-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   88.        N-(4-cyanophenyl)-3-[(4-fluorobenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   89.        3-[(4-fluorobenzoyl)amino]-N-fluorophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   90.        N-(4-chlorophenyl)-3-[(4-fluorobenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   91.        3-[(4-fluorobenzoyl)amino]-N-(4-methoxyphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   92.        3-[(4-fluorobenzoyl)amino]-N-(4-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   93.        N-(tert-butyl)-3-[(4-fluorobenzoyl)amino]-1H-thieno[2,3-c]pyrazolo-5-carboxamide;    -   94.        3-[(4-fluorobenzoyl)amino]-N-isopropyl-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   95.        N-ethyl-3[(4-flurobenzoyl)amino]-1N-thieno[2,3-c]pyrazole-5-carboxamide;    -   96.        4-chloro-N-{5-[(4-methylpiperazin-1-yl)carbonyl]-1H-thieno[2,3-e]pyrazol-3-yl}benzamide;    -   97.        4-chloro-N-[5-(morpholin-4-ylcarbonyl)-1H-thieno[2,3-c]pyrazol-3-yl]benzamide;    -   98.        3-[(4-chlorobenzoyl)amino]-N-(2-morpholinylethyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   99.        3-[(4-chlorobenzoyl)amino]-N-phenyl-1H-thieno[23]pyrazole-5-carboxamide;    -   100.        3-[(4-chlorobenzoyl)amino]-N-(2-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   101.        3-[(4-chlorobenzoyl)amino]-N-(4-cyanophenyl)-1H-thieno[2,3-c]pyrazole-5        carboxamide;    -   102.        3-[(4-chlorobenzoyl)amino]-N-fluorophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   103.        3-[(4-chlorobenzoyl)amino]N-(4-chlorophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   104.        3-[(4-chlorobenzoyl)amino]-N-(4-methoxyphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   105.        3-[(4-chlorobenzoyl)amino]-N-(4-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   106.        N-(tert-butyl)-3-[(4-chlorobenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   107.        3-[(4-chlorobenzoyl)amino]-N-isopropyl-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   108.        3-[(4-chlorobenzoyl)amino]-N-ethyl-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   109.        4-methoxy-N-{5-[(4-methylpiperazin-1-yl)carbonyl]-1H-thieno[2,3-c]pyrazol-3-yl}benzamide;    -   110.        4-methoxy-N-[5-(morpholin-4-ylcarbonyl)-1H-thieno[2,3-c]pyrazol-3-yl]benzamide;    -   111.        3-[(4-methoxybenzoyl)amino]-N-(2-morpholin-4-ylethyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   112.        3-[(4-methoxybenzoyl)amino]-N-phenyl-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   113.        3-[(4-methoxybenzoyl)amino]-N-(2-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   114.        N-(4-cyanophenyl)-3-[(4-methoxybenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   115.        N-(4-fluorophenyl)-3-[(4-methoxybenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   116.        N-(4-chlorophenyl)-3-[(4-methoxybenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   117.        3-[(4-methoxybenzoyl)amino]-N-(4-methoxyphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   118.        3-[(4-methoxybenzoyl)amino]-N-(4-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   119.        N-(tert-butyl)-3-[(4-methoxybenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   120.        N-isopropyl-3-[(4-methoxybenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   121.        N-ethyl-3-[(4-methoxybenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   122.        4-tert-butyl-N-{5-[(4-methylpiperazin-1-yl)carbonyl]-1H-thieno[2,3-c]pyrazol-3-yl}benzamide;    -   123.        4-tert-butyl-N-[5-(morpholin-4-ylcarbonyl)-1H-thieno[2,3-c]pyrazol-3-yl]benzamide;    -   124.        3-[(4-tert-butylbenzoyl)amino]-N-(2-morpholin-4-ylethyl)1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   125.        3-[(4-tert-butylbenzoyl)amino]-N-phenyl-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   126.        3-[(4-tert-butylbenzoyl)amino]-N-(2-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   127.        3-[(4-tert-butylbenzoyl)amino]-N-(4-cyanophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   128.        3-[(4-tert-butylbenzoyl)amino]-N-(4-fluorophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   129.        3-[(4-tert-butylbenzoyl)amino]-N-(4-chlorophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   130.    3-[(4-tert-butylbenzoyl)amino]-N-(4-methoxyphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   131.    3-[(4-tert-butylbenzoyl)amino]-N-(4-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   132.    N-(tert-butyl)-3-[(4-tert-butylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   133.    3-[(4-tert-butylbenzoyl)amino]-N-isopropyl-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   134.    3-[(4-tert-butylbenzoyl)amino]-N-ethyl-1H-thieno[2,3-c]pyrazol-5-carboxamide;

-   135.    4-methyl-N-{5-[(4-methylpiperazin-1-yl)carbonyl]-1H-thieno[2,3-c]pyrazol-3-yl}benzamide;

-   136.    4-methyl-N-[5-(morpholin-4-ylcarbonyl)-1H-thieno[2,3-c]pyrazol-3-yl]benzamide;

-   137.    3-[(4-methylbenzoyl)amino]-N-(2-morpholin-4-ylethyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   138.    3-[(4-methylbenzoyl)amino]-N-phenyl-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   139.    3-[(4-methylbenzoyl)amino]-N-(2-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   140.    N-(4-cyanophenyl)-3-[(4-methylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   141.    N-(4-fluorophenyl)-3-[(4-methylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   142.    N-(4-chlorophenyl)-3-[(4-methylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   143.    N-(4-methoxyphenyl)-3-[(4-methylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   144.    3-[(4-methylbenzoyl)amino]-N-(4-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   145.    N-(tert-butyl)-3-[(4-methylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   146.    N-isopropyl-3-[(4-methylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   147.    N-ethyl-3-[(4-methylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   148.    3-[(3-fluorobenzoyl)amino]-N-(4-morpholin-4-ylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   149.    N-(2-cyanophenyl)-3-[(3-fluorobenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   150.    3-[(3-fluorobenzoyl)amino]-N-(2-fluorophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   151.    N-(2-chlorophenyl)-3-[(3-fluorobenzoyl)amino]-1H-thieno[2,3-c]pyrazol    carboxamide;

-   152.    N-(2,6-dichlorophenyl)3-[(3-fluorobenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   153.    3-[(3-fluorobenzoyl)amino]-N-(2-methoxyphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   154.    N-(2,6-diethylphenyl)-3-[(3-fluorobenzoyl)amino]-1H-thieno[2,3-c]pyrazole-carboxamide;

-   155.    N-(3-cyanophenyl)-3-[(3-fluorobenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   156.    3-[(3-fluorobenzoyl)amino]-N-(3-fluorophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   157.    N-(3-chlorophenyl)-3-[(3-fluorobenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   158.    3-[(3-fluorobenzoyl)amino]-N-(3-methoxyphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   159.    3-[(3-fluorobenzoyl)amino]-N-(3-methylphenyl)-1H-thieno[2,3-e]pyrazole-5-carboxamide;

-   160.    3-[(3-fluorobenzoyl)amino]-N-[4-(4-methylpiperazin-1-yl)phenyl]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   161.    3-[(3-chlorobenzoyl)amino]-N-(4-morpholin-4-ylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   162.    3-[(3-chlorobenzoyl)amino]-N-(2-cyanophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   163.    3-[(3-chlorobenzoyl)amino]-N-2-fluorophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   164.    3-[(3-chlorobenzoyl)amino]-N-(2-chlorophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   165.    3-[(3-chlorobenzoyl)amino]-N-(2,6-dichlorophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   166.    3-[(3-chlorobenzoyl)amino]-N-(2-methoxyphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   167.    3-[(3-chlorobenzoyl)amino]-N-(2,6-diethylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   168.    3-[(3-chlorobenzoyl)amino]-N-(3-cyanophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   169.    3-[(3-chlorobenzoyl)amino]-N-(3-fluorophenyl)-1H-thieno[2,3-e]pyrazole-5-carboxamide;

-   170.    3-[(3-chlorobenzoyl)amino]-N-(3-chlorophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   171.    3-[(3-chlorobenzoyl)amino]-N-(3-methoxyphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   172.    3-[(3-chlorobenzoyl)amino]-N-(3-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   173.    3-[(3-chlorobenzoyl)amino]-N-[4-(4-methylpiperazin-1-yl)phenyl]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   174.    3-[(3-methoxybenzoyl)amino]-N-(4-morpholin-4-ylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   175.    N-2-cyanophenyl)-3-[(3-methoxybenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   176.    N-(2-fluorophenyl)-3-[(3-methoxybenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   177.    N-2-chlorophenyl)-3-[(3-methoxybenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   178.    N-(2,6-dichlorophenyl)-3-[(3-methoxybenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   179.    3-[(3-methoxybenzoyl)amino]-N-(2-methoxyphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   180.    N-(2,6-diethylphenyl)-3-[(3-methoxybenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   181.    N-(3-cyanophenyl)-3-[(3-methoxybenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   182.    N-(3-fluorophenyl)-3-[(3-methoxybenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   183.    N-(3-chlorophenyl)-3-[(3-methoxybenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   184.    3-[(3-methoxybenzoyl)amino]-N-(3-methoxyphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   185.    3-[(3-methoxybenzoyl)amino]-N-(3-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   186.    3-[(3-methoxybenzoyl)amino]-N-[4-(4-methylpiperazin-1-yl)phenyl]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   187.    N-(4-morpholin-4-ylphenyl)-3-[(phenylacetyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   188.    N-(2-cyanophenyl)-3-[(phenylacetyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   189.    N-(2-fluorophenyl)-3-[(phenylacetyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   190.    N-(2-chlorophenyl)-3-[(phenylacetyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   191.    N-(2,6-dichlorophenyl)-3-[(phenylacetyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

-   192.    N-(2-methoxyphenyl)-3-[(phenylacetyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   193.        N-(2,6-diethylphenyl)-3-[(phenylacetyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   194.        N-(3-cyanophenyl)-3-[(phenylacetyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   195.        N-(3-fluorophenyl)-3-[(phenylacetyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   196.        N-(3-chlorophenyl)-3-[(phenylacetyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   197.        N-(3-methoxyphenyl)-3-[(phenylacetyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   198.        N-(3-methylphenyl)-3-[(phenylacetyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   199.        N-[4-(4-methylpiperazin-1-yl)phenyl]-3-[(phenylacetyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   200.        3-{[(2-fluorophenyl)acetyl]amino}-N-(4-morpholin-4-ylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   201.        N-2-cyanophenyl)-3-{[(2-fluorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   202.        N-(2-fluorophenyl)-3-{[(2-fluorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   203.        N-(2-chlorophenyl)-3-{[(2-fluorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   204.        N-(2,6-dichlorophenyl)-3-{[(2-fluorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   205.        3-{[(2-fluorophenyl)acetyl]amino}-N-(2-methoxyphenyl)-1H-thieno[2,3-c]pyrazole-5        carboxamide;    -   206.        N-(2,6-diethylphenyl)-3-{[(2-fluorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   207. N-(3        cyanophenyl)-3-{[(2-fluorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   208.        N-(3-fluorophenyl)-3-{[(2-fluorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   209.        N-(3-chlorophenyl)-3-{[(2-fluorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   210.        3-{[(2-fluorophenyl)acetyl]amino}-N-(3-methoxyphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   211.        3-{[(2-fluorophenyl)acetyl]amino}-N-(3-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   212.        3-{[(2-fluorophenyl)acetyl]amino}-N-[4(4-methylpiperazin-1-yl)phenyl]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   213.        3-{[(2-methylphenyl)acetyl]amino}-N-(4-morpholin-4-ylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   214.        N-(2-cyanophenyl)-3-{[(2-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   215.        N-(2-fluorophenyl)-3-{[(2-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   216.        N-(2-chlorophenyl)-3-{[(2-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   217.        N-(2,6-dichlorophenyl)-3-{[(2-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   218.        N-(2-methoxyphenyl)-3-{[(2-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   219.        N-(2,6-diethylphenyl)-3-{[(2-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   220. N-(3        cyanophenyl)-3-{[(2-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   221.        N-(3-fluorophenyl)-3-{[(2-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   222.        N-(3-chlorophenyl)-3-{[(2-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   223.        N-(3-methoxyphenyl)-3-{[(2-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   224.        N-(3-methylphenyl)-3-{[(2-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   225.        3-{[(2-methylphenyl)acetyl]amino}-N-[4-(4-methylpiperazin-1-yl)phenyl]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   226.        3-{[(4-fluorophenyl)acetyl]amino}-N-(4-morpholin-4-ylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   227.        N-2-cyanophenyl)-3-{[(4-fluorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   228.        N-(2-fluorophenyl)-3-{[(4-fluorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   229.        N-(2-chlorophenyl)-3-{([(4-fluorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   230.        N-(2,6-dichlorophenyl)-3-{[(4-fluorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   231.        3-{[(4-fluorophenyl)acetyl]amino}-N-(2-methoxyphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   232.        N-(2,6-diethylphenyl)-3-{[(4-fluorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide    -   233.        N-(3-cyanophenyl)-3-{[(4-fluorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   234.        N-(3-fluorophenyl)-3-{[(4-fluorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   235.        N-(3-chlorophenyl)-3-{[(4-fluorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   236.        3-{[(4-fluorophenyl)acetyl]amino}-N-(3-methoxyphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   237.        3-{[(4-fluorophenyl)acetyl]amino}-N-(3-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   238.        3-{[(4-fluorophenyl)acetyl]amino}-N-[4-(4-methylpiperazin-1-yl)phenyl]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   239. 3-{[(4-chlorophenyl)acetyl]amino}-N-(4        morpholin-4-ylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   240.        3-{[(4-chlorophenyl)acetyl]amino}-N-(2-cyanophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   241.        3-{[(4-chlorophenyl)acetyl]amino}-N-(2-fluorophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   242.        N-2-chlorophenyl)-3-{[(4-chlorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   243.        3-{[(4-chlorophenyl)acetyl]amino}-N-(2,6-dichlorophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   244.        3-{[(4-chlorophenyl)acetyl]amino}-N-(2-methoxyphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   245.        3-{[(4-chlorophenyl)acetyl]amino}-N-(2,6-diethylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   246.        3-{[(4-chlorophenyl)acetyl]amino}-N-(3-cyanophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   247.        3-{[(4-chlorophenyl)acetyl]amino}-N-(3-fluorophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   248.        N-(3-chlorophenyl)-3-{[(4-chlorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   249.        3-{[(4-chlorophenyl)acetyl]amino}-N-(3-methoxyphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   250.        3-{[(4-chlorophenyl)acetyl]amino}-N-(3-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   251.        3-{[(4-chlorophenyl)acetyl]amino}-N-[4-(4-methylpiperazin-1-yl)phenyl]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   252.        3-{[(4-methoxyphenyl)acetyl]amino}-N-(4-morpholin-4-ylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   253.        N-(2-cyanophenyl)-3-{[(4-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   254.        N-(2-fluorophenyl)-3-{[(4-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   255.        N-(2-chlorophenyl)-3-{[(4-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   256.        N-(2,6-dichlorophenyl)-3-{[(4-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   257.        N-(2-methoxyphenyl)-3-{[(4-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   258.        N-(2,6-diethylphenyl)-3-{[(4-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   259.        N-(3-cyanophenyl)-3-{[(4-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   260.        N-(3-fluorophenyl)-3-{[(4-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   261.        N-(3-chlorophenyl)-3-{[(4-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   262.        N-(3-methoxyphenyl)-3-{[(4-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   263.        3-{[(4-methoxyphenyl)acetyl]amino}-N-(3-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   264.        3-{[(4-methoxyphenyl)acetyl]amino}-N-[4-(4-methylpiperazin-1-yl)phenyl]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   265.        3-{[(4-methylphenyl)acetyl]amino}-N-(4-morpholin-4-ylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   266.        N-(2-cyanophenyl)-3-{[(4-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   267.        N-(2-fluorophenyl)-3-{[(4-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   268.        N-(2-chlorophenyl)-3-{[(4-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   269.        N-(2,6-dichlorophenyl)-3-{[(4-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   270.        N-(2-methoxyphenyl-3-{[(4-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   271.        N-(2,6-diethylphenyl)-3-{[(4-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   272.        N-(3-cyanophenyl)-3-{[(4-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   273.        N-(3-fluorophenyl)-3-{[(4-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   274.        N-(3-chlorophenyl)-3-{[(4-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   275.        N-(3-methoxyphenyl)-3-{[(4-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   276.        N-(3-methylphenyl)-3-{[(4-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   277.        3-{[(4-methylphenyl)acetyl]amino}-N-[4-(4-methylpiperazin-1-yl)phenyl]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   278.        3-{[(4-tert-butylphenyl)acetyl]amino}-N-(4-morpholin-4-ylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   279.        3-{[(4-tert-butylphenyl)acetyl]amino}-N-(2-cyanophenyl)-1H-thieno[2,3-c]pyrazole-carboxamide;    -   280.        3-{[(4-tert-butylphenyl)acetyl]amino}-N-2-fluorophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   281.        3-{[(4-tert-butylphenyl)acetyl]amino}-N-(2-chlorophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   282.        3-{[(4-tert-butylphenyl)acetyl]amino}-N-(2,6-dichlorophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   283.        3-{[(4-tert-butylphenyl)acetyl]amino}-N-(2-methoxyphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   284.        3-{[(4-tert-butylphenyl)acetyl]amino}-N-(2,6-diethylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   285.        3-{[(4-tert-butylphenyl)acetyl]amino}-N-(3-cyanophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   286.        3-{[(4-tert-butylphenyl)acetyl]amino}-N-(3-fluorophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   287.        3-{[(4-tert-butylphenyl)acetyl]amino}-N-(3-chlorophenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   288. 3-{[(4        tert-butylphenyl)acetyl]amino}-N-(3-methoxyphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   289.        3-{[(4-tert-butylphenyl)acetyl]amino}-N-(3-methylphenyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   290.        3-{[(4-tert-butylphenyl)acetyl]amino}-N-[4-(4-methylpiperazin-1-yl)phenyl]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   291.        N-benzyl-3-{[(2-chlorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   292.        3-{[(2-chlorophenyl)acetyl]amino}-N-(2-fluorobenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   293.        N-2-chlorobenzyl)-3-{[(2-chlorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   294.        3-{[(2-chlorophenyl)acetyl]amino}-N-(2-methoxybenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   295.        3-{[(2-chlorophenyl)acetyl]amino}N-(2-methylbenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   296.        3-{[(2-chlorophenyl)acetyl]amino}-N-(3-fluorobenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   297.        3-{[(2-chlorophenyl)acetyl]amino}-N-(3-methoxybenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   298.        3-{[(2-chlorophenyl)acetyl]amino}-N-(3-methylbenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   299.        3-{[(2-chlorophenyl)acetyl]amino}-N-(4-fluorobenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   300.        N-(4-chlorobenzyl)-3-{[(2-chlorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   301.        3-{[(2-chlorophenyl)acetyl]amino}-N-(4-methoxybenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   302.        3-{([(2-chlorophenyl)acetyl]amino}-N-(4-methylbenzyl)-1H-thieno[2,3-c]pyrazol-5-carboxamide;    -   303.        N-(3-chlorobenzyl)-3-{[(2-chlorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   304.        N-benzyl-3-{[(2-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   305.        N-2-fluorobenzyl)-3-{[(2-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   306.        N-2-chlorobenzyl)-3-{[(2-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   307.        N-(2-methoxybenzyl)-3-{[(2-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   308.        3-{[(2-methoxyphenyl)acetyl]amino}-N-(2-methylbenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   309.        N-(3-fluorobenzyl)-3-{([(2-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   310.        N-(3-methoxybenzyl)-3-{[(2-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazol-5-carboxamide;    -   311.        3-{[(2-methoxyphenyl)acetyl]amino}-N-(3-methylbenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   312.        N-(4-fluorobenzyl)-3-{[(2-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   313.        N-(4-chlorobenzyl)-3-{[(2-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5        carboxamide;    -   314.        N-(4-methoxybenzyl)-3-{[(2-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   315.        3-{[(2-methoxyphenyl)acetyl]amino}-N-(4-methylbenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   316.        N-(3-chlorobenzyl)-3-{[(2-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   317.        N-benzyl-3-{[(3-fluorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   318.        N-2-fluorobenzyl)-3-{[(3-fluorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   319.        N-2-chlorobenzyl)-3-{[(3-fluorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   320.        3-{[(3-fluorophenyl)acetyl]amino}-N-(2-methoxybenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   321.        3-{[(3-fluorophenyl)acetyl]amino}-N-(2-methylbenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   322.        N-(3-fluorobenzyl)-3-{[(3-fluorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   323.        3-{[(3-fluorophenyl)acetyl]amino}-N-(3-methoxybenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   324.        3-{[(3-fluorophenyl)acetyl]amino}-N-(3-methylbenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   325.        N-(4-fluorobenzyl)-3-{[(3-fluorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   326.        N-(4-chlorobenzyl)-3-{[(3-fluorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   327.        3-{[(3-fluorophenyl)acetyl]amino}-N-(4-methoxybenzyl)-1H-thieno[2,3-c]pyrazole-5        carboxamide;    -   328.        3-{[(3-fluorophenyl)acetyl]amino}-N-(4-methylbenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   329.        N-(3-chlorobenzyl)-3-{[(3-fluorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide,    -   330.        N-benzyl-3-{[(3-chlorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   331.        3-{[(3-chlorophenyl)acetyl]amino}-N-2-fluorobenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   332.        N-(2-chlorobenzyl)-3-{[(3-chlorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   333.        3-{[(3-chlorophenyl)acetyl]amino}-N-(2-methoxybenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   334. 3-{[(3        chlorophenyl)acetyl]amino}-N-(2-methylbenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   335.        3-{[(3-chlorophenyl)acetyl]amino}-N-(3-fluorobenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   336.        3-{[(3-chlorophenyl)acetyl]amino}-N-(3-methoxybenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   337.        3-{[3-chlorophenyl)acetyl]amino}-N-(3-methylbenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   338.        3-{[(3-chlorophenyl)acetyl]amino})-N-fluorobenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   339.        N-(4-chlorobenzyl)-3-{[(3-chlorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   340.        3-{[(3-chlorophenyl)acetyl]amino}-N-(4-methoxybenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   341.        3-{[(3-chlorophenyl)acetyl]amino}-N-(4-methylbenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   342.        N-(3-chlorobenzyl)-3-{[(3-chlorophenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   343.        N-benzyl-3-{[(3-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   344.        N-(2-fluorobenzyl)-3-{[(3-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   345.        N-(2-chlorobenzyl)-3-{[(3-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   346.        N-(2-methoxybenzyl)-3-{[(3-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   347.        3-{[(3-methoxyphenyl)acetyl]amino}-N-(2-methylbenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   348.        N-(3-fluorobenzyl)-3-{[(3-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   349.        N-(3-methoxybenzyl)-3-{[(3-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   350.        3-{[(3-methoxyphenyl)acetyl]amino}-N-(3-methylbenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   351.        N-(4-fluorobenzyl)-3-{[(3-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   352.        N-(4-chlorobenzyl)-3-{[(3-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   353.        N-(4-methoxybenzyl)-3-{[(3-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   354.        3-{[(3-methoxyphenyl)acetyl]amino}-N-(4-methylbenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   355.        N-(3-chlorobenzyl)-3-{[(3-methoxyphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   356.        N-benzyl-3-{[(3-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   357.        N-2-fluorobenzyl)-3-{[(3-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   358.        N-(2-chlorobenzyl)-3-{[(3-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   359.        N-(2-methoxybenzyl)-3-{[(3-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   360.        N-(2-methylbenzyl)-3-{[(3-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   361.        N-(3-fluorobenzyl)-3-{[(3-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   362.        N-(3-methoxybenzyl)-3-{[(3-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   363.        N-(3-methylbenzyl)-3-{[(3-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   364.        N-(4-fluorobenzyl)-3-{[(3-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazol-5-carboxamide;    -   365.        N-(chlorobenzyl)-3-{[(3-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5        carboxamide;    -   366. N-(4        methoxybenzyl)-3-{[(3-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   367.        N-(4-methylbenzyl)-3-{[(3-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   368.        N-(3-chlorobenzyl)-3-{[(3-methylphenyl)acetyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide,    -   369.        N-benzyl-3-[(thien-2-ylcarbonyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   370.        N-(2-fluorobenzyl)-3-[(thien-2-ylcarbonyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   371.        N-2-chlorobenzyl)-3-[(thien-2-ylcarbonyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   372.        N-(2-methoxybenzyl)-3-[(thien-2-ylcarbonyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   373.        N-(2-methylbenzyl)-3-[(thien-2-ylcarbonyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   374.        N-(3-fluorobenzyl)-3-[(thien-2-ylcarbonyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   375.        N-(3-methoxybenzyl)-3-[(thien-2-ylcarbonyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   376.        N-(3-methylbenzyl)-3-[(thien-2-ylcarbonyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   377.        N-(4-fluorobenzyl)-3-[(thien-2-ylcarbonyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   378.        N-(4-chlorobenzyl)-3-[(thien-2-ylcarbonyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   379.        N-(4-methoxybenzyl)-3-[(thien-2-ylcarbonyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   380.        N-(4-methylbenzyl)-3-[(thien-2-ylcarbonyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   381.        N-(3-chlorobenzyl)-3-[(thien-2-ylcarbonyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   382.        N-benzyl-3-(isonicotinoylamino)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   383.        N-(2-fluorobenzyl)-3-(isonicotinoylamino)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   384.        N-(2-chlorobenzyl)-3-(isonicotinoylamino)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   385.        3-(isonicotinoylamino)-N-(2-methoxybenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   386.        3-isonicotinoylamino)-N-(2-methylbenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   387.        N-(3-fluorobenzyl)-3-(isonicotinoylamino)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   388.        3-(isonicotinoylamino)-N-(3-methoxybenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   389.        3-(isonicotinoylamino)-N-(3-methylbenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   390.        N-(4-fluorobenzyl)-3-(isonicotinoylamino)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   391. N-(4        chlorobenzyl)-3-(isonicotinoylamino)-1H-thieno[2,3]pyrazole-5-carboxamide;    -   392.        3-isonicotinoylamino)-N-(4-methoxybenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   393.        3-(isonicotinoylamino)-N-(4-methylbenzyl)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   394.        N-(3-chlorobenzyl)-3-(isonicotinoylamino)-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   395.        N-benzyl-3-[(1,3-thiazol-2-ylcarbonyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   396.        N-(2-fluorobenzyl)-3-[(1,3-thiazol-2-ylcarbonyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   397.        N-(2-chlorobenzyl)-3-[(1,3-thiazol-2-ylcarbonyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   398.        N-(2-methoxybenzyl)-3-[(1,3-thiazol-2-ylcarbonyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   399.        N-(2-methylbenzyl)-3-[(1,3-thiazol-2-ylcarbonyl)amino]-1H-thieno[2,3-c]pyrazole-5        carboxamide;    -   400.        N-(3-fluorobenzyl)-3-[(1,3-thiazol-2-ylcarbonyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   401.        N-(3-methoxybenzyl)-3-[(1,3-thiazol-2-ylcarbonyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   402.        N-(3-methylbenzyl)-3-[(1,3-thiazol-2-ylcarbonyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   403.        N-(4-fluorobenzyl)-3-[(1,3-thiazol-2-ylcarbonyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   404.        N-(4-chlorobenzyl)-3-[(1,3-thiazol-2-ylcarbonyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   405.        N-(4-methoxybenzyl)-3-[(1,3-thiazol-2-ylcarbonyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   406.        N4-methylbenzyl)-3-[(1,3-thiazol-2-ylcarbonyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   407.        N-(3-chlorobenzyl)-3-[(1,3-thiazol-2-ylcarbonyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   408.        N-benzyl-3-{[4-(4-methylpiperazin-1-yl)benzoyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   409.        N-2-fluorobenzyl)-3-{[4-(4-methylpiperazin-1-yl)benzoyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   410.        N-(2-chlorobenzyl)-3-{[4-(4-methylpiperazin-1-yl)benzoyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   411.        N-(2-methoxybenzyl)-3-{[4-(4-methylpiperazin-1-yl)benzoyl]amino}-1H-thieno[2,3-c]pyrazole-5        carboxamide;    -   412.        N-(2-methylbenzyl)-3-{[4-(4-methylpiperazin-1-yl)benzoyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   413.        N-(3-fluorobenzyl)-3-{[4-(4-methylpiperazin-1-yl)benzoyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   414.        N-(3-methoxybenzyl)-3-{[4-(4-methylpiperazin-1-yl)benzoyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   415.        N-(3-methylbenzyl)-3-{[4-(4-methylpiperazin-1-yl)benzoyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   416.        N-(4-fluorobenzyl)-3-{[4-(4-methylpiperazin-1-yl)benzoyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   417.        N-(4-chlorobenzyl)-3-{[4-(4-methylpiperazin-1-yl)benzoyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   418.        N-(4-methoxybenzyl)-3-{[4-(4-methylpiperazin-1-yl)benzoyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   419.        N-(4-methylbenzyl)-3-{[4-(4-methylpiperazin-1-yl)benzoyl]amino}-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   420.        N-(3-chlorobenzyl)-3-{[4(4-methylpiperazin-1-yl)benzoyl]amino}-1H-thieno[2,3-c]pyrazole-5        carboxamide;    -   421.        N-benzyl-3-[(4-morpholin-4-ylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   422.        N-2-fluorobenzyl)-3-[(4-morpholin-4-ylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5        carboxamide;    -   423.        N2-chlorobenzyl)-3-[(4-morpholin-4-ylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   424.        N-(2-methoxybenzyl)-3-[(4-morpholin-4-ylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   425.        N-(2-methylbenzyl)-3-[(4-morpholin-4-ylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   426.        N-(3-fluorobenzyl)-3-[(4-morpholin-4-ylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   427.        N-(3-methoxybenzyl)-3-[(4-morpholin-4-ylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   428.        N-(3-methylbenzyl)-3-[(4-morpholin-4-ylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   429.        N-(4-fluorobenzyl)-3-[(4-morpholin-4-ylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   430.        N-(4-chlorobenzyl)-3-[(4-morpholin-4-ylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   431.        N-(4-methoxybenzyl)-3-[(4-morpholin-4-ylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   432.        N-(4-methylbenzyl)-3-[(4-morpholin-4-ylbenzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;    -   433.        N-(3-chlorobenzyl)-3-[(4-morpholin-4-benzoyl)amino]-1H-thieno[2,3-c]pyrazole-5-carboxamide;

1. A combinatorial chemical library comprising a plurality of members offormula (VIII)

wherein X is NR′, O, S, SO or SO₂; each of R and R₁, being the same ordifferent, is independently a hydrogen atom or an optionally substitutedgroup selected from R′, —COR′, —COOR′, —CONHR′, —CONR′R″, —SO₂R′,—SO₂NHR′ or —SO₂NR′R″; wherein each of R′ and R″, being the same ordifferent, is independently selected from hydrogen or an optionallyfurther substituted straight or branched C₁-C₆ alkyl, aryl or aryl-C₁-C₆alkyl group; R₂ is an optionally substituted group selected from R′,—CH₂OR′ and OR′, wherein R′ is as above defined; and thepharmaceutically acceptable salts thereof.
 2. The library of claim 1wherein the resin is a polystyrenic resin.
 3. A chemical library ofcompounds comprising two or more derivatives of formula (I)

wherein X is NR′, O, S, SO or SO₂; each of R and R₁, being the same ordifferent, is independently a hydrogen atom or an optionally substitutedgroup selected from R′, —COR′, —COOR′, —CONHR′, —CONR′R″, —SO₂R′,—SO₂NHR′ or —SO₂NR′R″; wherein each of R′ and R″, being the same ordifferent, is independently selected from hydrogen or an optionallyfurther substituted straight or branched C₁-C₆ alkyl, aryl or aryl-C₁-C₆alkyl group; R₂ is an optionally substituted group selected from R′,—CH₂OR′ and OR′, wherein R′ is as above defined; and thepharmaceutically acceptable salts thereof with the proviso that suchderivatives are not4-phenyl-5-(phenylsulfonyl)-2H-thieno[2,3-c]pyrazol-3-amine or5-phenyl-1H-furo[2,3-c]pyrazol-3-amine.
 4. The polystyrenic resinaccording to claim 2 wherein said resin is methylisocyanate polystyrenicresin.